Abstract
Lentiviral vectors are now recognised as an efficient transgene delivery system which can result in greater than 90% of founder animals carrying the transgene. Vector injection into the perivitelline space has emerged as the standard delivery method but is limited by the need for high-titre lentivirus vector preparations. Based on a modified perivitelline injection method we demonstrate that transgenic animals can be generated from low-titre virus vector preparations further simplifying lentiviral transgenesis. Repeat injection of 107 TU/ml vector preparation resulted in 23% of embryos carrying the transgene compare to 1% from a single injection. Embryos exposed to repeat injection of vector developed to blastocyst with the same efficiency as non-injected embryos and produced transgenic mice capable of transmitting the transgene through the germline
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References
Breslow NE, Clayton DG (1993) Approximate inference in generalized linear mixed models. J Am Stat Assoc 88(421):9–25
Ewerling S, Hofmann A, Klose R, Weppert M, Brem G, Rink K, Pfeifer A, Wolf E (2006) Evaluation of laser-assisted lentiviral transgenesis in bovine. Transgenic Res 15(4):447–454
Green DP (1997) Three-dimensional structure of the zona pellucida. Rev Reprod 2(3):147–156
Hofmann A, Kessler B, Ewerling S, Weppert W, Vogg B, Ludwig H, Stojkovic M, Boelhauve M, Brem G, Wolf E, Pfeifer A (2003) Efficient transgenesis in farm animals by lentiviral vectors. EMBO Rep 4(11):1054–1060
Hofmann A, Zakhartchenko V, Weppert M, Sebald H, Wenigerkind H, Brem G, Wolf E, Pfeifer A (2004) Generation of transgenic cattle by lentiviral gene transfer into oocytes. Biol Reprod 71(2):405–409
Lois C, Hong EJ, Pease S, Brown EJ, Baltimore D (2002) Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295(5556):868–872
McGrew MJ, Sherman A, Ellard FM, Lillico SG, Gilhooley HJ, Kingsman AJ, Mitrophanous KA, Sang H (2004) Efficient production of germline transgenic chickens using lentiviral vectors. EMBO Rep 5(7):728–733
Pfeifer A (2004) Lentiviral transgenesis. Transgenic Res 13(6):513–522
Pfeifer A, Ikawa M, Dayn Y, Verma IM (2002) Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proc Natl Acad Sci USA 99(4):2140–2145
Ritchie WA, Taylor JE, Gardner JO, Wilmut I, Carlisle A, Neil C, King T, Whitelaw CBA (2005) Live lambs born from zona-pellucida denuded embryos. Cloning Stem Cells 7(3):178–182
Whitelaw CBA, Radcliffe PA, Ritchie WA, Carlisle A, Ellard FM, Pena RN, Rowe J, Clark AJ, King TJ, Mitrophanous KA (2004) Efficient generation of transgenic pigs using equine infectious anaemia virus (EIAV) derived vector. FEBS Lett 571(1–3):233–236
Acknowledgements
We are grateful to the animal staff for care of the animals and to Caroline McCorquodale for statistical analysis. The pgk-GFP lentivirus vector was supplied by Genethon (France). This work was supported by the BBSRC and through the EC NEST029025 project INTEGRA.
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Ritchie, W.A., Neil, C., King, T. et al. Transgenic embryos and mice produced from low titre lentiviral vectors. Transgenic Res 16, 661–664 (2007). https://doi.org/10.1007/s11248-007-9102-2
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DOI: https://doi.org/10.1007/s11248-007-9102-2