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Efficient generation of transgenic mice by direct intraovarian injection of plasmid DNA.

Abstract：

:We established a rapid procedure for obtaining transgenic mice by directly injecting an enhanced green fluorescent protein (EGFP)-expressing plasmid (pIRES-EGFP) into the ovaries of fertile mice. The frequency of transgenic mouse production was determined by pair-mating, and by polymerase chain reaction (PCR) and sequence analysis of DNA taken from the tails of the offspring. The mice that received the EGFP gene transmitted it to their offspring (F(1)). Genetic and PCR analyses of F(1) progeny confirmed that the inserted EGFP was stably inherited. Of six female F(1) mice, all were able to pass the foreign DNA on to the next generation (F(2)). In situ hybridization using paraffin-embedded sections of ovarian and testicular tissues from the F(1) and F(2) progeny showed that the introduced gene was expressed in the gonads of the animals. The chromosomal location of the injected DNA was determined by fluorescence in situ hybridization, and the frequency of multiple site versus single site insertions is 85.71% (18/21) analyzed by FISH. We anticipate great progress in murine genetic engineering using this technique.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Yang SY,Wang JG,Cui HX,Sun SG,Li Q,Gu L,Hong Y,Liu PP,Liu WQ

doi

10.1016/j.bbrc.2007.04.112

subject

Has Abstract

pub_date

2007-06-22 00:00:00

pages

266-71

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(07)00842-X

journal_volume

358

pub_type

杂志文章

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