Transgenic embryos and mice produced from low titre lentiviral vectors.
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 10(7) 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.
Ritchie
William A
WA
Animal Services Department, Roslin Institute, Roslin, Midlothian EH25 9PS, UK.
Neil
Clare
C
King
Tim
T
Whitelaw
C Bruce A
CB
D000818:Animals; D030801:Animals, Genetically Modified; D001709:Biotechnology / Q000379:methods; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes