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Transfection

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230: 607:. This inflammation hypersensitizes the exposed cell and nearby cells to subsequent exposure. As a result, while a cell can be repeatedly transfected with short RNA with few non-specific effects, repeatedly transfecting cells with even a small amount of long RNA can cause cell death unless measures are taken to suppress or evade the innate immune system (see "Long-RNA transfection" below). 2880: 170:, magnetofection); others rely on chemical materials or biological particles (viruses) that are used as carriers. There are many different methods of gene delivery developed for various types of cells and tissues, from bacterial to mammalian. Generally, the methods can be divided into three categories: physical, chemical, and biological. 720:), and others. All of these proteins can specifically bind to exogenous RNA molecules and trigger an immune response. The specific chemical, structural or other characteristics of long RNA molecules that are required for recognition by PRRs remain largely unknown despite intense study. At any given time, a typical 149:
into cells, resulting in an infection. For work with bacterial and archaeal cells transfection retains its original meaning as a special case of transformation. Because the term transformation had another sense in animal cell biology (a genetic change allowing long-term propagation in culture, or
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Protoplast fusion is a technique in which transformed bacterial cells are treated with lysozyme in order to remove the cell wall. Following this, fusogenic agents (e.g., Sendai virus, PEG, electroporation) are used in order to fuse the protoplast carrying the gene of interest with the target
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solution containing the DNA to be transfected. When the two are combined, a fine precipitate of the positively charged calcium and the negatively charged phosphate will form, binding the DNA to be transfected on its surface. The suspension of the precipitate is then added to the cells to be
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Menuel S, Fontanay S, Clarot I, Duval RE, Diez L, Marsura A (December 2008). "Synthesis and complexation ability of a novel bis- (guanidinium)-tetrakis-(beta-cyclodextrin) dendrimeric tetrapod as a potential gene delivery (DNA and siRNA) system. Study of cellular siRNA transfection".
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from the cell death caused by frequent transfection with long, protein-encoding RNA. Inhibiting interferon signaling disrupts the positive-feedback loop that normally hypersensitizes cells exposed to exogenous long RNA. Researchers have recently used this technique to express
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For some applications of transfection, it is sufficient if the transfected genetic material is only transiently expressed. Since the DNA introduced in the transfection process is usually not integrated into the nuclear genome, the foreign DNA will be diluted through
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vectors can be useful for viral transfection methods because they can transfer genes into a wide variety of human cells and have high transfer rates. Lentiviral vectors are also helpful due to their ability to transduce cells not currently undergoing mitosis.
534:" of proteins of interests (e.g. Endothelin-1) with potential applications in gene therapy. Limitation of the silencing approach are the toxicity of the transfection for cells and potential "off-target" effects on the expression of other genes/proteins. 740:, and specifically that 5'-triphosphate RNA, which can be produced during viral infection, is more immunogenic than 5'-diphosphate RNA, 5'-monophosphate RNA or RNA containing no 5' phosphate. However, in vitro-transcribed (ivT) long RNA containing a 1204:
Fischer D, von Harpe A, Kunath K, Petersen H, Li Y, Kissel T (2002). "Copolymers of ethylene imine and N-(2-hydroxyethyl)-ethylene imine as tools to study effects of polymer structure on physicochemical and biological properties of DNA complexes".
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transfected (usually a cell culture grown in a monolayer). By a process not entirely understood, the cells take up some of the precipitate, and with it, the DNA. This process has been a preferred method of identifying many oncogenes.
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Long-RNA transfection is the process of deliberately introducing RNA molecules longer than about 25nt into living cells. A distinction is made between short- and long-RNA transfection because exogenous long RNA molecules elicit an
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is a method used in mice and rats, in which nucleic acids can be delivered to the liver by injecting a relatively large volume in the blood in less than 10 seconds; nearly all of the DNA is expressed in the liver by this procedure.
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Arnold AS, Laporte V, Dumont S, Appert-Collin A, Erbacher P, Coupin G, Levy R, Poindron P, Gies JP (February 2006). "Comparing reagents for efficient transfection of human primary myoblasts: FuGENE 6, Effectene and ExGen 500".
603:, which is triggered by longer RNA molecules. Most cells of the body express proteins of the innate immune system, and upon exposure to exogenous long RNA molecules, these proteins initiate signaling cascades that result in 462:(EBV) nuclear antigen 1 (EBNA1) or the SV40 large-T antigen allow episomal amplification of plasmids containing the viral EBV (293E) or SV40 (293T) origins of replication, greatly reducing the rate of dilution. 1490: 630:) that encode a short RNA molecule can also be used, short-RNA transfection does not risk modification of the cell's DNA, a characteristic that has led to the development of short RNA as a new class of 766:
nucleotides. Although only a very small number of these modified nucleotides are present in a typical mRNA molecule, they may help prevent mRNA from activating the innate immune system by disrupting
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Electroporator with square wave and exponential decay waveforms for in vitro, in vivo, adherent cell and 96 well electroporation applications. Manufactured by BTX Harvard Apparatus, Holliston MA USA.
197:, which is a lipid-mediated DNA-transfection process utilizing liposome vectors. It can also include the use of polymeric gene carriers (polyplexes). Biological transfection is typically mediated by 937: 241:, where short electrical pulses disrupt the cell membrane, allowing the transfected nucleic acids to enter the cell. Other physical methods use different means to poke holes in the cell membrane: 201:, utilizing the ability of a virus to inject its DNA inside a host cell. A gene that is intended for delivery is packaged into a replication-deficient viral particle. Viruses used to date include 770:
that would resemble double-stranded RNA (dsRNA), a type of RNA thought to be present in cells only during viral infection. The immunogenicity of long RNA has been used to study both innate and
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Yoneyama M, Kikuchi M, Natsukawa T, Shinobu N, Imaizumi T, et al. (2004). "The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses".
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acquisition of properties typical of cancer cells), the term transfection acquired, for animal cells, its present meaning of a change in cell properties caused by introduction of DNA.
477:, while other cells will die. After applying this selective stress (selection pressure) for some time, only the cells with a stable transfection remain and can be cultivated further. 1011:
Saul JM, Linnes MP, Ratner BD, Giachelli CM, Pun SH (November 2007). "Delivery of non-viral gene carriers from sphere-templated fibrin scaffolds for sustained transgene expression".
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Sapra, Rachit; Verma, Ram P.; Maurya, Govind P.; Dhawan, Sameer; Babu, Jisha; Haridas, V. (13 November 2019). "Designer Peptide and Protein Dendrimers: A Cross-Sectional Analysis".
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mRNA) is also highly immunogenic despite having no 5' phosphate, suggesting that characteristics other than 5'-phosphorylation can influence the immunogenicity of an RNA molecule.
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Physical methods are the conceptually simplest, using some physical means to force the transfected material into the target cell's nucleus. The most widely used physical method is
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is a series of widely used proprietary non-liposomal transfection reagents capable of directly transfecting a wide variety of cells with high efficiency and low toxicity.
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The meaning of the term has evolved. The original meaning of transfection was "infection by transformation", i.e., introduction of genetic material, DNA or RNA, from a
2264:"Protein synthesis in rabbit reticulocytes. Purification and characterization of a double-stranded RNA-dependent protein synthesis inhibitor from reticulocyte lysates" 465:
If it is desired that the transfected gene actually remain in the genome of the cell and its daughter cells, a stable transfection must occur. To accomplish this, a
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Stable and transient transfection differ in their long term effects on a cell; a stably transfected cell will continuously express transfected DNA and pass it on to
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can also be transfected to achieve RNA silencing (i.e. loss of RNA and protein from the targeted gene). This has become a major application in research to achieve "
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cells, including plant cells. In animal cells, transfection is the preferred term as transformation is also used to refer to progression to a cancerous state (
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Heitz, Marc; Javor, Sacha; Darbre, Tamis; Reymond, Jean-Louis (21 August 2019). "Stereoselective pH Responsive Peptide Dendrimers for siRNA Transfection".
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Alexopoulou L, Holt AC, Medzhitov R, Flavell RA (2001). "Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3".
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recipient cell. A major disadvantage of this method is that bacterial components are non-specifically introduced into the target cell as well.
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is a class of highly branched molecules based on various building blocks and synthesized through a convergent or a divergent method. These
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molecules. How cells distinguish exogenous long RNA from the large amount of endogenous long RNA is an important open question in
2123:"Suppression of RNA recognition by Toll-like receptors: the impact of nucleoside modification and the evolutionary origin of RNA" 1092:
Mehier-Humbert S, Guy RH (April 2005). "Physical methods for gene transfer: improving the kinetics of gene delivery into cells".
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Short-RNA transfection is routinely used in biological research to knock down the expression of a protein of interest (using
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Tsukakoshi M, Kurata S, Nomiya Y, et al. (1984). "A Novel Method of DNA Transfection by Laser Microbeam Cell Surgery".
264:, in which nucleic acid is attached to heavy metal particles (usually gold) and propelled into the cells at high speed; and 1957: 3808: 3803: 3549: 3465: 3451: 3121: 1307:"Transfer of the gene for thymidine kinase to thymidine kinase-deficient human cells by purified herpes simplex viral DNA" 800: 3305: 3212: 3090: 678: 3813: 3439: 3424: 3368: 3359: 3310: 3235: 3176: 381: 261: 3913: 3470: 3376: 3271: 3143: 3134: 2923: 1985: 846: 3923: 3665: 3607: 3511: 3446: 3429: 3340: 3194: 3038: 2994: 1270:
Graham FL, van der Eb AJ (April 1973). "A new technique for the assay of infectivity of human adenovirus 5 DNA".
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Felgner JH, Kumar R, Sridhar CN, Wheeler CJ, Tsai YJ, Border R, Ramsey P, Martin M, Felgner PL (January 1994).
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Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (November 1987).
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Jacobsen LB, Calvin SA, Colvin KE, Wright M (June 2004). "FuGENE 6 Transfection Reagent: the gentle power".
836: 426: 65: 459: 3928: 3918: 771: 574: 430: 294:, polymers, liposomes, or nanoparticles (with or without chemical or viral functionalization. See below). 214: 2305:"Characterization of purified double-stranded RNA-activated eIF-2 alpha kinase from rabbit reticulocytes" 3220: 3018: 2984: 2748: 1899: 646: 611: 469:
is co-transfected, which gives the cell some selectable advantage, such as resistance towards a certain
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RNA can also be transfected into cells to transiently express its coded protein, or to study
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Hellgren I, Drvota V, Pieper R, Enoksson S, Blomberg P, Islam KB, Sylvén C (August 2000).
733: 562: 392:. The efficiency of lipofection can be improved by treating transfected cells with a mild 389: 303: 238: 174: 116: 104: 2164:"Innate antiviral responses by means of TLR7-mediated recognition of single-stranded RNA" 2903: 2885: 2782: 2668: 2557: 2418: 2361: 2179: 2039: 1591: 1566: 1406: 1322: 1059: 870: 3712: 3707: 3642: 3544: 3381: 3345: 2687: 2652: 2625: 2600: 2576: 2541: 2435: 2402: 1962: 1024: 988: 963: 816: 737: 558: 546: 531: 307: 265: 257: 178: 163: 61: 41: 2321: 2304: 2280: 2263: 1466: 1449: 1425: 1390: 1341: 1306: 557:
template. As with DNA, RNA can be delivered to cells by a variety of means including
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Transfection can result in unexpected morphologies and abnormalities in target cells.
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transfection) is a technique used to inject genetic material into a cell by means of
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RNA molecules shorter than about 25nt (nucleotides) largely evade detection by the
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machinery, and therefore is not referred to as siRNA). While DNA-based vectors (
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Proceedings of the National Academy of Sciences of the United States of America
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Proceedings of the National Academy of Sciences of the United States of America
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kinetics. RNA transfection is often used in primary cells that do not divide.
409: 393: 335: 314:-buffered saline solution (HeBS) containing phosphate ions is combined with a 269: 246: 210: 206: 202: 142: 68:
is often used to describe virus-mediated gene transfer into eukaryotic cells.
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the RNA into the encoded protein. If the RNA is a regulatory RNA (such as a
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Several methods use tools that force the nucleic acid into the cell, namely:
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Takahasi K, Yoneyama M, Nishihori T, Hirai R, Kumeta H, et al. (2008).
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Das HK, Das A, Ghosh-Dastidar P, Ralston RO, Yaghmai B, et al. (1981).
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Pipes BL, Vasanwala FH, Tsang TC, Zhang T, Luo P, Harris DT (January 2005).
1415: 1391:"Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure" 1129:
Nonviral Vectors for Gene Therapy - Physical Methods and Medical Translation
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Schlee M, Roth A, Hornung V, Hagmann CA, Wimmenauer V, et al. (2009).
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bind the nucleic acids to form dendriplexes that then penetrate the cells.
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Biology Research Resource — Articles and Forums about Transfection
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to allow the uptake of material. Transfection can be carried out using
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Schmidt A, Schwerd T, Hamm W, Hellmuth JC, Cui S, et al. (2009).
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Luo D, Saltzman WM (January 2000). "Synthetic DNA delivery systems".
2501:"Structural mechanism of RNA recognition by the RIG-I-like receptors" 2344:
Hornung V, Ellegast J, Kim S, Brzozka K, Jung A, et al. (2006).
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Segura T, Shea LD (2001). "Materials for non-viral gene delivery".
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Primary and Stem Cells: Gene Transfer Technologies and Applications
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in cells that can cause a variety of nonspecific effects including
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Herb M, Farid A, Gluschko A, Krönke M, Schramm M (November 2019).
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Herb M, Farid A, Gluschko A, Krönke M, Schramm M (November 2019).
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Eukaryotic mRNA contains chemically modified nucleotides such as
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uses a highly focused laser to form a ~1 μm diameter hole.
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that fuse with the cell membrane and deposit their cargo inside.
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Research in optical transfection at the University of St Andrews
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Saito T; Owen DM; Jiang F; Marcotrigiano J; Gale M, Jr. (2008).
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Diebold SS, Kaisho T, Hemmi H, Akira S, Reis e Sousa C (2004).
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or mixtures) to form an aggregate with the negatively charged (
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typically involves opening transient pores or "holes" in the
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is the process of deliberately introducing naked or purified
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Kariko K, Ni H, Capodici J, Lamphier M, Weissman D (2004).
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cell may contain several hundred thousand mRNA and other,
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Process of introducing nucleic acids into eukaryotic cells
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uses high-intensity ultrasound (attributed mainly to the
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The innate immune system has evolved to protect against
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of gas bubbles interacting with nearby cell membranes),
2079:"mRNA is an endogenous ligand for Toll-like receptor 3" 1131:. Advances in Genetics. Vol. 89. pp. 89–111. 736:
of the 5'-end of a long RNA molecule can influence its
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particles and driven into the target cells by magnets.
1958:"How nanotechnology helps mRNA Covid-19 vaccines work" 480:
Common agents for selecting stable transfection are:
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as a carrier. In such cases, the technique is called
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List of varieties of genetically modified maize/corn
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The 10th US-Japan Symposium on Drug Delivery Systems
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Applied Physics B: Photophysics and Laser Chemistry
368:. Lipofection generally uses a positively charged ( 1904:: CS1 maint: DOI inactive as of September 2024 ( 158:There are various methods of introducing foreign 99:constructs), may be transfected. Transfection of 2121:Kariko K, Buckstein M, Ni H, Weissman D (2005). 1127:Suda T, Liu D (2015). "Hydrodynamic Delivery". 334:(PEI). The negatively charged DNA binds to the 268:, where nucleic acids are attached to magnetic 1795:Durocher Y, Perret S, Kamen A (January 2002). 887: 885: 883: 487:, or G418, neutralized by the product of the 3098: 2944: 2346:"5'-triphosphate RNA is the ligand for RIG-I" 545:either chemically, or enzymatically using an 8: 1846:"The Science of Stable Cell Line Generation" 1366:Transfer and Expression: A Laboratory Manual 421:DNA can also be introduced into cells using 338:and the complex is taken up by the cell via 3740:Detection of genetically modified organisms 2749:"RNA offers a safer way to reprogram cells" 2116: 2114: 1984:Foley, Katherine Ellen (22 December 2020). 614:) or to express or block the activity of a 429:, and the cells are said to be transduced. 3597: 3507: 3364: 3139: 3130: 3105: 3091: 3083: 2951: 2937: 2929: 2646: 2644: 1241:"Nanoparticle Based Transfection Reagents" 48:" is typically used to describe non-viral 2906:at the U.S. National Library of Medicine 2852: 2723: 2686: 2676: 2624: 2575: 2565: 2516: 2475: 2434: 2369: 2320: 2303:Levin DH, Petryshyn R, London IM (1981). 2279: 2187: 2138: 2094: 1932: 1820: 1590: 1506: 1465: 1424: 1414: 1340: 1330: 987: 957: 955: 953: 951: 924:at the U.S. National Library of Medicine 873:at the U.S. National Library of Medicine 592:was a breakthrough for producing viable 1652:Fundamental & Clinical Pharmacology 863: 458:or degraded. Cell lines expressing the 1897: 1622:Lakshmipathy U, Thyagarajan B (2011). 1245:Biology Transfection Research Resource 968:Analytical and Bioanalytical Chemistry 671:pathogen-associated molecular patterns 1956:Cooney, Elizabeth (1 December 2020). 1305:Bacchetti S, Graham FL (April 1977). 537:RNA can be purified from cells after 7: 1571:Cellular and Molecular Life Sciences 260:of nucleic acid with a fine needle; 119:, by cell squeezing, or by mixing a 2770:Annual Review of Materials Research 1534:. Transfection of Mammalian Cells. 1454:The Journal of Biological Chemistry 962:Kim TK, Eberwine JH (August 2010). 681:(PRRs) for exogenous RNA including 290:can be divided into several kinds: 3770:Genetic use restriction technology 1866:Mawji IA, Marsden PA (June 2006). 1025:10.1016/j.biomaterials.2007.07.026 588:Encapsulating the RNA molecule in 25: 2716:Journal of Visualized Experiments 1925:Journal of Visualized Experiments 1872:Experimental Biology and Medicine 1628:(1st ed.). Wiley-Blackwell. 1368:. W. H. Freeman. pp. 96–97. 1247:. Transfection.ws. Archived from 661:Endogenous vs. exogenous long RNA 442:Stable and transient transfection 298:One of the cheapest methods uses 2878: 1664:10.1111/j.1472-8206.2005.00344.x 897:Protocols and Applications Guide 782:Inhibiting only three proteins, 732:. Several reports suggest that 677:". Many cells express specific 3788:Cartagena Protocol on Biosafety 1758:10.1021/acs.bioconjchem.9b00403 360:that can easily merge with the 2791:10.1146/annurev.matsci.31.1.25 1886:(inactive 12 September 2024). 794:is sufficient to rescue human 778:Repeated long-RNA transfection 364:since they are both made of a 1: 3550:Somatic cell nuclear transfer 2499:Yoneyama M, Fujita T (2008). 2322:10.1016/S0021-9258(19)69008-3 2281:10.1016/S0021-9258(19)69192-1 1467:10.1016/S0021-9258(17)41980-6 679:pattern recognition receptors 322:Another method is the use of 126:with the material to produce 2678:10.1371/journal.pone.0011756 2617:10.1016/j.immuni.2009.05.008 2518:10.1016/j.immuni.2008.07.009 2477:10.1016/j.molcel.2007.11.028 2140:10.1016/j.immuni.2005.06.008 1884:10.3181/00379727-231-2310704 1284:10.1016/0042-6822(73)90341-3 1137:10.1016/bs.adgen.2014.10.002 943:Dorland's Medical Dictionary 2011:Tansey B (11 August 2006). 1707:10.1021/acs.chemrev.9b00153 1544:10.1016/j.ymeth.2003.11.002 567:lipid-mediated transfection 302:, originally discovered by 288:Chemical-based transfection 262:biolistic particle delivery 3945: 2747:Trafton A (26 July 2010). 2651:Angel M, Yanik MF (2010). 2015:. San Francisco Chronicle. 1106:10.1016/j.addr.2004.12.007 847:Vector (molecular biology) 3873: 3608:Genetically modified food 3039:Sister chromatid exchange 980:10.1007/s00216-010-3821-6 899:. Promega. Archived from 541:or synthesized from free 173:Physical methods include 3034:Horizontal gene transfer 2964:homologous recombination 2908:Medical Subject Headings 926:Medical Subject Headings 875:Medical Subject Headings 573:, transfected cells may 569:. If the RNA encodes a 2968:mobile genetic elements 2567:10.1073/pnas.0900971106 2371:10.1126/science.1132505 2189:10.1126/science.1093616 1416:10.1073/pnas.84.21.7413 583:RNAi-mediated knockdown 93:supercoiled plasmid DNA 2546:Proc Natl Acad Sci USA 2096:10.1074/jbc.M310175200 1801:Nucleic Acids Research 1746:Bioconjugate Chemistry 1332:10.1073/pnas.74.4.1590 1207:Bioconjugate Chemistry 1171:Bioconjugate Chemistry 801:reprogramming proteins 643:innate immune response 234: 215:adeno-associated virus 3221:Roundup ready soybean 3019:Chromosomal crossover 2854:10.1038/nmeth1105-875 922:Transduction, Genetic 742:7-methylguanosine cap 276:Hydrodynamic delivery 232: 3760:Reverse transfection 3535:Genetic transduction 2800:Nature Biotechnology 601:innate immune system 366:phospholipid bilayer 310:in 1973 (see also). 251:optical transfection 219:herpes simplex virus 191:hydrostatic pressure 145:-infecting virus or 113:tricalcium phosphate 3750:Genetics in fiction 3682:Genetic enhancement 3484:Hepatitis B vaccine 3114:Genetic engineering 2783:2001AnRMS..31...25S 2669:2010PLoSO...511756A 2558:2009PNAS..10612067S 2552:(29): 12067–12072. 2427:10.1038/nature07106 2419:2008Natur.454..523S 2362:2006Sci...314..964H 2180:2004Sci...303.1529D 2174:(5663): 1529–1531. 2089:(13): 12542–12550. 2040:2001Natur.413..732A 1813:10.1093/nar/30.2.e9 1701:(21): 11391–11441. 1407:1987PNAS...84.7413F 1364:Kriegler M (1991). 1323:1977PNAS...74.1590B 1060:1984ApPhB..35..135T 768:secondary structure 726:regulatory long RNA 590:lipid nanoparticles 3860:Stem cell research 3479:Ice-minus bacteria 2888:has a profile for 2835:Bonetta L (2005). 2751:. MIT News Office. 1844:Fanelli A (2016). 1583:10.1007/PL00000769 1094:Adv Drug Deliv Rev 1068:10.1007/BF00697702 683:toll-like receptor 460:Epstein–Barr virus 374:cationic liposomes 235: 162:into a eukaryotic 91:material (such as 64:) in these cells. 3914:Molecular biology 3901: 3900: 3865:Synthetic biology 3755:Human enhancement 3745:Genetic pollution 3721: 3720: 3589: 3588: 3497: 3496: 3460: 3459: 3354: 3353: 3080: 3079: 2894: 2413:(7203): 523–527. 2356:(5801): 994–997. 2315:(14): 7638–7641. 2274:(12): 6491–6495. 2034:(6857): 732–738. 1635:978-0-470-61074-9 1508:10.2144/05381bm05 1375:978-0-7167-7004-6 1219:10.1021/bc025550w 1183:10.1021/bc800193p 803:in primary human 772:adaptive immunity 386:calcium phosphate 324:cationic polymers 300:calcium phosphate 109:calcium phosphate 16:(Redirected from 3936: 3924:Applied genetics 3638:Dow AgroSciences 3598: 3508: 3365: 3140: 3131: 3107: 3100: 3093: 3084: 2953: 2946: 2939: 2930: 2892: 2882: 2881: 2866: 2856: 2831: 2794: 2753: 2752: 2744: 2738: 2737: 2727: 2707: 2701: 2700: 2690: 2680: 2648: 2639: 2638: 2628: 2596: 2590: 2589: 2579: 2569: 2537: 2531: 2530: 2520: 2496: 2490: 2489: 2479: 2455: 2449: 2448: 2438: 2398: 2392: 2391: 2373: 2341: 2335: 2334: 2324: 2300: 2294: 2293: 2283: 2259: 2253: 2252: 2216: 2210: 2209: 2191: 2159: 2153: 2152: 2142: 2118: 2109: 2108: 2098: 2074: 2068: 2067: 2048:10.1038/35099560 2023: 2017: 2016: 2008: 2002: 2001: 1999: 1997: 1981: 1975: 1974: 1972: 1970: 1953: 1947: 1946: 1936: 1916: 1910: 1909: 1903: 1895: 1863: 1857: 1856: 1854: 1852: 1841: 1835: 1834: 1824: 1792: 1786: 1785: 1752:(8): 2165–2182. 1741: 1735: 1734: 1695:Chemical Reviews 1690: 1684: 1683: 1646: 1640: 1639: 1619: 1613: 1612: 1594: 1577:(8–9): 1326–33. 1562: 1556: 1555: 1527: 1521: 1520: 1510: 1486: 1480: 1479: 1469: 1445: 1439: 1438: 1428: 1418: 1386: 1380: 1379: 1361: 1355: 1354: 1344: 1334: 1302: 1296: 1295: 1267: 1261: 1260: 1258: 1256: 1251:on 21 April 2013 1237: 1231: 1230: 1201: 1195: 1194: 1165: 1159: 1158: 1124: 1118: 1117: 1089: 1080: 1079: 1043: 1037: 1036: 1008: 1002: 1001: 991: 959: 946: 935: 929: 919: 913: 912: 910: 908: 889: 878: 868: 760:5-methylcytidine 756:-methyladenosine 706:protein kinase R 516:RNA transfection 332:polyethylenimine 316:calcium chloride 308:A. J. van der Eb 283:Chemical methods 225:Physical methods 21: 3944: 3943: 3939: 3938: 3937: 3935: 3934: 3933: 3904: 3903: 3902: 3897: 3869: 3838: 3792: 3774: 3727: 3717: 3691: 3687:Genetic testing 3669: 3659: 3585: 3571:Recombinant DNA 3559: 3530:Electroporation 3493: 3489:Oncolytic virus 3468: 3456: 3413: 3399:Herman the Bull 3350: 3284: 3266: 3248: 3230: 3207: 3189: 3171: 3125: 3123: 3116: 3111: 3081: 3076: 3062:Antigenic shift 3048: 3024:Gene conversion 2999: 2970: 2957: 2900: 2899: 2898: 2883: 2879: 2874: 2869: 2847:(11): 875–883. 2834: 2797: 2766: 2762: 2760:Further reading 2757: 2756: 2746: 2745: 2741: 2709: 2708: 2704: 2650: 2649: 2642: 2598: 2597: 2593: 2539: 2538: 2534: 2498: 2497: 2493: 2457: 2456: 2452: 2400: 2399: 2395: 2343: 2342: 2338: 2302: 2301: 2297: 2261: 2260: 2256: 2218: 2217: 2213: 2161: 2160: 2156: 2120: 2119: 2112: 2076: 2075: 2071: 2025: 2024: 2020: 2010: 2009: 2005: 1995: 1993: 1983: 1982: 1978: 1968: 1966: 1955: 1954: 1950: 1918: 1917: 1913: 1896: 1865: 1864: 1860: 1850: 1848: 1843: 1842: 1838: 1794: 1793: 1789: 1743: 1742: 1738: 1692: 1691: 1687: 1648: 1647: 1643: 1636: 1621: 1620: 1616: 1564: 1563: 1559: 1529: 1528: 1524: 1488: 1487: 1483: 1447: 1446: 1442: 1388: 1387: 1383: 1376: 1363: 1362: 1358: 1304: 1303: 1299: 1269: 1268: 1264: 1254: 1252: 1239: 1238: 1234: 1203: 1202: 1198: 1177:(12): 2357–62. 1167: 1166: 1162: 1147: 1126: 1125: 1121: 1091: 1090: 1083: 1045: 1044: 1040: 1019:(31): 4705–16. 1010: 1009: 1005: 961: 960: 949: 936: 932: 920: 916: 906: 904: 903:on 25 June 2014 891: 890: 881: 869: 865: 860: 813: 780: 764:2'-O-methylated 734:phosphorylation 663: 563:electroporation 518: 491:resistance gene 444: 419: 390:electroporation 285: 239:electroporation 227: 175:electroporation 156: 139: 117:electroporation 56:and non-animal 28: 23: 22: 15: 12: 11: 5: 3942: 3940: 3932: 3931: 3926: 3921: 3916: 3906: 3905: 3899: 3898: 3896: 3895: 3890: 3885: 3880: 3874: 3871: 3870: 3868: 3867: 3862: 3857: 3852: 3846: 3844: 3843:Similar fields 3840: 3839: 3837: 3836: 3831: 3826: 3816: 3811: 3806: 3800: 3798: 3794: 3793: 3791: 3790: 3784: 3782: 3776: 3775: 3773: 3772: 3767: 3762: 3757: 3752: 3747: 3742: 3737: 3731: 3729: 3723: 3722: 3719: 3718: 3716: 3715: 3713:Gene targeting 3710: 3708:Gene knockdown 3705: 3699: 3697: 3693: 3692: 3690: 3689: 3684: 3679: 3673: 3671: 3661: 3660: 3658: 3657: 3656: 3655: 3650: 3645: 3643:DuPont Pioneer 3640: 3635: 3630: 3622: 3617: 3616: 3615: 3604: 3602: 3601:In agriculture 3595: 3591: 3590: 3587: 3586: 3584: 3583: 3578: 3573: 3567: 3565: 3561: 3560: 3558: 3557: 3552: 3547: 3545:Microinjection 3542: 3537: 3532: 3527: 3522: 3516: 3514: 3505: 3499: 3498: 3495: 3494: 3492: 3491: 3486: 3481: 3475: 3473: 3462: 3461: 3458: 3457: 3455: 3454: 3449: 3444: 3443: 3442: 3437: 3427: 3421: 3419: 3415: 3414: 3412: 3411: 3406: 3401: 3396: 3391: 3390: 3389: 3384: 3382:Knockout mouse 3373: 3371: 3362: 3356: 3355: 3352: 3351: 3349: 3348: 3343: 3338: 3333: 3328: 3323: 3318: 3313: 3308: 3303: 3298: 3292: 3290: 3286: 3285: 3283: 3282: 3276: 3274: 3268: 3267: 3265: 3264: 3258: 3256: 3250: 3249: 3247: 3246: 3240: 3238: 3232: 3231: 3229: 3228: 3223: 3217: 3215: 3209: 3208: 3206: 3205: 3199: 3197: 3191: 3190: 3188: 3187: 3181: 3179: 3173: 3172: 3170: 3169: 3164: 3159: 3154: 3148: 3146: 3137: 3128: 3118: 3117: 3112: 3110: 3109: 3102: 3095: 3087: 3078: 3077: 3075: 3074: 3069: 3064: 3058: 3056: 3050: 3049: 3047: 3046: 3041: 3036: 3031: 3026: 3021: 3016: 3010: 3008: 3001: 3000: 2998: 2997: 2995:Transformation 2992: 2987: 2981: 2979: 2972: 2971: 2958: 2956: 2955: 2948: 2941: 2933: 2927: 2926: 2921: 2916: 2911: 2884: 2877: 2876: 2875: 2873: 2872:External links 2870: 2868: 2867: 2841:Nature Methods 2832: 2795: 2763: 2761: 2758: 2755: 2754: 2739: 2702: 2640: 2591: 2532: 2511:(2): 178–181. 2491: 2470:(4): 428–440. 2450: 2393: 2336: 2295: 2254: 2233:10.1038/ni1087 2227:(7): 730–737. 2211: 2154: 2133:(2): 165–175. 2110: 2069: 2018: 2003: 1992:. Quartz Media 1976: 1948: 1911: 1878:(6): 704–708. 1858: 1836: 1787: 1736: 1685: 1641: 1634: 1614: 1557: 1522: 1481: 1460:(4): 2550–61. 1440: 1401:(21): 7413–7. 1381: 1374: 1356: 1297: 1262: 1232: 1213:(5): 1124–33. 1196: 1160: 1145: 1119: 1081: 1054:(3): 135–140. 1038: 1003: 947: 938:"Transfection" 930: 914: 893:"Transfection" 879: 862: 861: 859: 856: 855: 854: 849: 844: 839: 834: 832:Transformation 829: 824: 819: 817:Gene targeting 812: 809: 779: 776: 738:immunogenicity 702:RIG1 (RARRES3) 662: 659: 632:macromolecular 559:microinjection 547:RNA polymerase 517: 514: 513: 512: 507: 502: 497: 492: 448:daughter cells 443: 440: 418: 415: 414: 413: 403: 397: 343: 320: 284: 281: 266:magnetofection 258:microinjection 226: 223: 179:microinjection 155: 152: 138: 135: 62:carcinogenesis 46:transformation 26: 24: 14: 13: 10: 9: 6: 4: 3: 2: 3941: 3930: 3929:Biotechnology 3927: 3925: 3922: 3920: 3919:Gene delivery 3917: 3915: 3912: 3911: 3909: 3894: 3891: 3889: 3888:Biotechnology 3886: 3884: 3881: 3879: 3876: 3875: 3872: 3866: 3863: 3861: 3858: 3856: 3853: 3851: 3848: 3847: 3845: 3841: 3835: 3832: 3830: 3829:South America 3827: 3824: 3820: 3819:North America 3817: 3815: 3812: 3810: 3807: 3805: 3802: 3801: 3799: 3795: 3789: 3786: 3785: 3783: 3781: 3777: 3771: 3768: 3766: 3765:Transhumanism 3763: 3761: 3758: 3756: 3753: 3751: 3748: 3746: 3743: 3741: 3738: 3736: 3733: 3732: 3730: 3724: 3714: 3711: 3709: 3706: 3704: 3703:Gene knockout 3701: 3700: 3698: 3694: 3688: 3685: 3683: 3680: 3678: 3675: 3674: 3672: 3667: 3662: 3654: 3651: 3649: 3646: 3644: 3641: 3639: 3636: 3634: 3631: 3629: 3626: 3625: 3623: 3621: 3618: 3614: 3613:Controversies 3611: 3610: 3609: 3606: 3605: 3603: 3599: 3596: 3592: 3582: 3579: 3577: 3574: 3572: 3569: 3568: 3566: 3562: 3556: 3553: 3551: 3548: 3546: 3543: 3541: 3538: 3536: 3533: 3531: 3528: 3526: 3523: 3521: 3518: 3517: 3515: 3513: 3512:Inserting DNA 3509: 3506: 3504: 3500: 3490: 3487: 3485: 3482: 3480: 3477: 3476: 3474: 3472: 3467: 3463: 3453: 3450: 3448: 3445: 3441: 3438: 3436: 3433: 3432: 3431: 3428: 3426: 3423: 3422: 3420: 3418:Other animals 3416: 3410: 3407: 3405: 3402: 3400: 3397: 3395: 3392: 3388: 3385: 3383: 3380: 3379: 3378: 3375: 3374: 3372: 3370: 3366: 3363: 3361: 3357: 3347: 3344: 3342: 3339: 3337: 3334: 3332: 3329: 3327: 3324: 3322: 3319: 3317: 3314: 3312: 3309: 3307: 3304: 3302: 3299: 3297: 3294: 3293: 3291: 3287: 3281: 3278: 3277: 3275: 3273: 3269: 3263: 3260: 3259: 3257: 3255: 3251: 3245: 3242: 3241: 3239: 3237: 3233: 3227: 3224: 3222: 3219: 3218: 3216: 3214: 3210: 3204: 3201: 3200: 3198: 3196: 3192: 3186: 3183: 3182: 3180: 3178: 3174: 3168: 3165: 3163: 3160: 3158: 3155: 3153: 3150: 3149: 3147: 3145: 3141: 3138: 3136: 3132: 3129: 3127: 3119: 3115: 3108: 3103: 3101: 3096: 3094: 3089: 3088: 3085: 3073: 3070: 3068: 3065: 3063: 3060: 3059: 3057: 3055: 3051: 3045: 3042: 3040: 3037: 3035: 3032: 3030: 3027: 3025: 3022: 3020: 3017: 3015: 3012: 3011: 3009: 3007: 3002: 2996: 2993: 2991: 2988: 2986: 2983: 2982: 2980: 2978: 2973: 2969: 2965: 2961: 2954: 2949: 2947: 2942: 2940: 2935: 2934: 2931: 2925: 2922: 2920: 2917: 2915: 2912: 2909: 2905: 2902: 2901: 2896: 2895: 2891:transfection 2887: 2871: 2864: 2860: 2855: 2850: 2846: 2842: 2838: 2833: 2829: 2825: 2821: 2817: 2813: 2812:10.1038/71889 2809: 2805: 2801: 2796: 2792: 2788: 2784: 2780: 2776: 2772: 2771: 2765: 2764: 2759: 2750: 2743: 2740: 2735: 2731: 2726: 2725:10.3791/60143 2721: 2717: 2713: 2706: 2703: 2698: 2694: 2689: 2684: 2679: 2674: 2670: 2666: 2663:(7): e11756. 2662: 2658: 2654: 2647: 2645: 2641: 2636: 2632: 2627: 2622: 2618: 2614: 2610: 2606: 2602: 2595: 2592: 2587: 2583: 2578: 2573: 2568: 2563: 2559: 2555: 2551: 2547: 2543: 2536: 2533: 2528: 2524: 2519: 2514: 2510: 2506: 2502: 2495: 2492: 2487: 2483: 2478: 2473: 2469: 2465: 2461: 2454: 2451: 2446: 2442: 2437: 2432: 2428: 2424: 2420: 2416: 2412: 2408: 2404: 2397: 2394: 2389: 2385: 2381: 2377: 2372: 2367: 2363: 2359: 2355: 2351: 2347: 2340: 2337: 2332: 2328: 2323: 2318: 2314: 2310: 2306: 2299: 2296: 2291: 2287: 2282: 2277: 2273: 2269: 2265: 2258: 2255: 2250: 2246: 2242: 2238: 2234: 2230: 2226: 2222: 2215: 2212: 2207: 2203: 2199: 2195: 2190: 2185: 2181: 2177: 2173: 2169: 2165: 2158: 2155: 2150: 2146: 2141: 2136: 2132: 2128: 2124: 2117: 2115: 2111: 2106: 2102: 2097: 2092: 2088: 2084: 2080: 2073: 2070: 2065: 2061: 2057: 2053: 2049: 2045: 2041: 2037: 2033: 2029: 2022: 2019: 2014: 2007: 2004: 1991: 1987: 1980: 1977: 1965: 1964: 1959: 1952: 1949: 1944: 1940: 1935: 1934:10.3791/60143 1930: 1926: 1922: 1915: 1912: 1907: 1901: 1893: 1889: 1885: 1881: 1877: 1873: 1869: 1862: 1859: 1847: 1840: 1837: 1832: 1828: 1823: 1818: 1814: 1810: 1806: 1802: 1798: 1791: 1788: 1783: 1779: 1775: 1771: 1767: 1763: 1759: 1755: 1751: 1747: 1740: 1737: 1732: 1728: 1724: 1720: 1716: 1712: 1708: 1704: 1700: 1696: 1689: 1686: 1681: 1677: 1673: 1669: 1665: 1661: 1657: 1653: 1645: 1642: 1637: 1631: 1627: 1626: 1618: 1615: 1610: 1606: 1602: 1598: 1593: 1588: 1584: 1580: 1576: 1572: 1568: 1561: 1558: 1553: 1549: 1545: 1541: 1538:(2): 104–12. 1537: 1533: 1526: 1523: 1518: 1514: 1509: 1504: 1500: 1496: 1495:BioTechniques 1492: 1485: 1482: 1477: 1473: 1468: 1463: 1459: 1455: 1451: 1444: 1441: 1436: 1432: 1427: 1422: 1417: 1412: 1408: 1404: 1400: 1396: 1392: 1385: 1382: 1377: 1371: 1367: 1360: 1357: 1352: 1348: 1343: 1338: 1333: 1328: 1324: 1320: 1317:(4): 1590–4. 1316: 1312: 1308: 1301: 1298: 1293: 1289: 1285: 1281: 1278:(2): 456–67. 1277: 1273: 1266: 1263: 1250: 1246: 1242: 1236: 1233: 1228: 1224: 1220: 1216: 1212: 1208: 1200: 1197: 1192: 1188: 1184: 1180: 1176: 1172: 1164: 1161: 1156: 1152: 1148: 1146:9780128022726 1142: 1138: 1134: 1130: 1123: 1120: 1115: 1111: 1107: 1103: 1100:(5): 733–53. 1099: 1095: 1088: 1086: 1082: 1077: 1073: 1069: 1065: 1061: 1057: 1053: 1049: 1042: 1039: 1034: 1030: 1026: 1022: 1018: 1014: 1007: 1004: 999: 995: 990: 985: 981: 977: 974:(8): 3173–8. 973: 969: 965: 958: 956: 954: 952: 948: 945: 944: 939: 934: 931: 927: 923: 918: 915: 902: 898: 894: 888: 886: 884: 880: 876: 872: 867: 864: 857: 853: 850: 848: 845: 843: 840: 838: 835: 833: 830: 828: 825: 823: 820: 818: 815: 814: 810: 808: 806: 802: 797: 793: 789: 785: 777: 775: 773: 769: 765: 761: 757: 755: 749: 747: 743: 739: 735: 731: 727: 723: 719: 715: 711: 707: 703: 700: 696: 692: 688: 684: 680: 676: 672: 669:by detecting 668: 660: 658: 656: 652: 648: 644: 638: 636: 633: 629: 625: 621: 617: 613: 608: 606: 602: 597: 595: 591: 586: 584: 580: 576: 572: 568: 564: 560: 556: 552: 548: 544: 540: 535: 533: 529: 525: 523: 515: 511: 510:Blasticidin S 508: 506: 503: 501: 498: 496: 493: 490: 486: 483: 482: 481: 478: 476: 472: 468: 463: 461: 457: 451: 449: 441: 439: 435: 432: 428: 424: 417:Viral methods 416: 411: 407: 404: 401: 398: 395: 391: 387: 383: 379: 375: 371: 367: 363: 362:cell membrane 359: 355: 351: 347: 344: 341: 337: 333: 329: 325: 321: 317: 313: 309: 305: 301: 297: 296: 295: 293: 289: 282: 280: 277: 273: 271: 267: 263: 259: 254: 252: 248: 244: 240: 231: 224: 222: 220: 216: 212: 208: 204: 200: 196: 192: 188: 187:impalefection 184: 180: 176: 171: 169: 168:nanoparticles 165: 161: 153: 151: 148: 147:bacteriophage 144: 136: 134: 131: 129: 125: 122: 118: 114: 110: 106: 105:cell membrane 102: 98: 94: 90: 86: 82: 78: 74: 69: 67: 63: 59: 55: 51: 47: 43: 40: 36: 35:nucleic acids 32: 19: 3677:Gene therapy 3576:Transgenesis 3555:Transfection 3554: 3520:Agrobacteria 3404:Knockout rat 3226:Vistive Gold 3122:Genetically 3067:Reassortment 3014:Transfection 3013: 2990:Transduction 2904:Transfection 2890: 2844: 2840: 2803: 2799: 2774: 2768: 2742: 2715: 2705: 2660: 2656: 2611:(1): 25–34. 2608: 2604: 2594: 2549: 2545: 2535: 2508: 2504: 2494: 2467: 2463: 2453: 2410: 2406: 2396: 2353: 2349: 2339: 2312: 2308: 2298: 2271: 2267: 2257: 2224: 2220: 2214: 2171: 2167: 2157: 2130: 2126: 2086: 2082: 2072: 2031: 2027: 2021: 2006: 1994:. Retrieved 1989: 1979: 1967:. Retrieved 1961: 1951: 1924: 1914: 1900:cite journal 1875: 1871: 1861: 1849:. Retrieved 1839: 1804: 1800: 1790: 1749: 1745: 1739: 1698: 1694: 1688: 1655: 1651: 1644: 1624: 1617: 1574: 1570: 1560: 1535: 1531: 1525: 1501:(1): 48–52. 1498: 1494: 1484: 1457: 1453: 1443: 1398: 1394: 1384: 1365: 1359: 1314: 1310: 1300: 1275: 1271: 1265: 1255:30 September 1253:. Retrieved 1249:the original 1244: 1235: 1210: 1206: 1199: 1174: 1170: 1163: 1128: 1122: 1097: 1093: 1051: 1047: 1041: 1016: 1013:Biomaterials 1012: 1006: 971: 967: 941: 933: 917: 905:. Retrieved 901:the original 896: 871:Transfection 866: 852:Viral vector 837:Transduction 827:Protofection 784:interferon-β 781: 753: 750: 744:(present in 730:cell biology 675:inflammation 664: 653:arrest, and 639: 609: 605:inflammation 598: 594:RNA vaccines 587: 536: 526: 519: 505:Hygromycin B 479: 464: 452: 445: 436: 427:transduction 420: 405: 399: 382:DEAE-dextran 356:, which are 345: 328:DEAE-dextran 323: 304:F. L. 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Index

Lipofection
nucleic acids
eukaryotic
cells
transformation
DNA
bacteria
eukaryotic
carcinogenesis
Transduction
portmanteau
Genetic
supercoiled plasmid DNA
siRNA
animal cells
cell membrane
calcium phosphate
tricalcium phosphate
electroporation
cationic
lipid
liposomes
prokaryote
bacteriophage
DNA
cell
nanoparticles
electroporation
microinjection
gene gun

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