[Generation of transgenic mice by intratesticular injection and electroporation in vivo].
OBJECTIVE
To evaluate the feasibility of establishing transgenic mice by means of seminiferous tubule microinjection and electroporation (EP) in vivo.


METHODS
Specific pathogen-free (SPF) male Kunming mice divided into 4 groups were subjected to microinjection of two different transfection solutions labeled with enhance green fluorescent protein (EGFP) into the seminiferous tubule of the testis, and in one of the two groups receiving the identical transfection solutions, EP in vivo was performed. After two weeks, the male mice of each group were mated with SPF female Kunming mice with superovulation treatment, and PCR coupled with Southern blotting was performed for the offspring mice.


RESULTS
The results of PCR suggested significant difference in the efficiency of exogenous gene integration between the 4 groups (P<0.01), among which group A achieved the greatest efficiency (45%). Southern blotting did not identify significant difference between the 4 groups (P>0.05), but still suggested the highest efficiency in group A (25%).


CONCLUSIONS
Seminiferous tubule microinjection in conjunction with subsequent EP in vivo can remarkably enhance the integration efficiency of exogenous genes into the host genome, but this new method needs to be further tested for its potential utility in transgenic animal generation.
Yuan
Jin
J
Center of Laboratory Animals, Southern Medical University, Guangzhou 510515, China. yuanjin@fimmu.com
An
Jing
J
Gu
Wei-Wang
WW
Huang
Wu-Jian
WJ
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections; D016133:Polymerase Chain Reaction; D012671:Seminiferous Tubules; D014162:Transfection / Q000379:methods
D049452:Green Fluorescent Proteins; D004247:DNA