Selection of mouse preimplantation embryos carrying exogenous DNA by polymerase chain reaction.
A polymerase chain reaction (PCR) system to detect transgenes in mouse preimplantation embryos was employed so that transgenic embryos could be selected before they were transferred to recipient mice. The selection system involves bisection of morulae, selection of the half-morulae containing target sequences within 7 hr, and culture and transfer of the sister half-morulae. PCR analysis of morulae derived from transgenic mice confirmed that the PCR system was reliable. However, five of 41 implanted embryos derived from PCR-positive morulae did not contain the transgenes. Also, one of 28 implanted embryos from PCR-negative morulae were transgenic. The selection system was applied to fertilized mouse eggs into which pSV2-gpt-gE1A DNA was injected. The injected DNA was detected in 30 of 84 morulae derived from the microinjected eggs. All seven implanted embryos developed from PCR-negative morulae had no detectable amount of transgenes, and one of two successfully implanted embryos from PCR-positive morulae was transgenic.
Ninomiya
T
T
Research Institute of Life Science, Snow Brand Milk Products, Co., Ltd., Tochigi, Japan.
Hoshi
M
M
Mizuno
A
A
Nagao
M
M
Yuki
A
A
D000818:Animals; D002970:Cleavage Stage, Ovum / Q000032:analysis*; D004247:DNA / Q000032:analysis*; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D008845:Microinjections; D009028:Morula / Q000032:analysis*; D016133:Polymerase Chain Reaction / Q000379:methods; D013698:Templates, Genetic
D004247:DNA