Gene transfer efficiency during gestation and the influence of co-transfer of non-manipulated embryos on production of transgenic mice.
Litter size of DNA microinjected zygotes is lower than for non-manipulated zygotes. The rate of embryonic and fetal survival in early, mid and late gestation was determined to assess whether DNA integration was responsible for embryonic losses. Also, the effect of including non-microinjected embryos with injected embryos on pregnancy rate and transgenic pup production was determined. In Experiment 1, one-cell embryos from immature CD-1 mice were microinjected with a whey acidic protein promoter-human protein C gene construct. One hour after microinjection embryos were transferred to pseudopregnant recipients (45 transfers of 30 embryos each). Fifteen recipients were sacrificed on day 4, 12 and 18 of gestation and the embryos/fetuses analysed for the transgene. The percentage of embryos or fetuses that were positive for the transgene was not significantly different at any day. However, the number of viable embryos at day 4 was significantly greater than fetuses on days 12 or 18. In addition, a high degree of mosaicism was observed in day 18 fetuses and placentae recovered. In Experiment 2, one-cell embryos from CD-1 mice were microinjected and co-transferred with non-manipulated embryos (C57BL/6). Pregnancy rate and the total number of pups born were improved by addition of non-injected embryos. However, the number of transgenic mice produced was similar whether non-injected embryos were included or not. There were 32.2% (15/46) transgenic pups when 0 non-injected embryos were transferred compared with 15.1% (13/86) transgenic pups when 4 or 8 non-injected embryos were added to the transfers. In summary, a high degree of embryonic and fetal mortality occurs among microinjected embryos.(ABSTRACT TRUNCATED AT 250 WORDS)
Canseco
R S
RS
Department of Dairy Science, Virginia Polytechnic Institute and State University, Blacksburg 24061-0315.
Sparks
A E
AE
Page
R L
RL
Russell
C G
CG
Johnson
J L
JL
Velander
W H
WH
Pearson
R E
RE
Drohan
W N
WN
Gwazdauskas
F C
FC
D000818:Animals; D000831:Animals, Newborn / Q000235:genetics / Q000502:physiology; D004247:DNA / Q000235:genetics*; D004624:Embryo Transfer / Q000662:veterinary*; D005260:Female; D005313:Fetal Death / Q000662:veterinary; D005865:Gestational Age; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic / Q000196:embryology* / Q000235:genetics*; D011247:Pregnancy; D014162:Transfection / Q000235:genetics
D004247:DNA