Germ line transmission and expression of a corrected HPRT gene produced by gene targeting in embryonic stem cells.
The deletion mutation in the HPRT-deficient mouse embryonic stem (ES) cell line E14TG2a has been corrected by gene targeting. The presence of plasmid sequences in the correcting vector DNA did not affect the frequency of correction. We have characterized three different HPRT gene structures in correctants. Cells from one corrected clone have been introduced into mouse blastocysts, and germ line transmission of the ES cell-derived corrected gene has been achieved. The corrected gene has the same pattern of expression as the wild-type gene, with the characteristic elevated level of expression in brain tissue. Hence, we have demonstrated the feasibility of introducing targeted modifications into the mouse germ line by homologous recombination in ES cells.
Thompson
S
S
Department of Molecular Biology, University of Edinburgh, Scotland.
Clarke
A R
AR
Pow
A M
AM
Hooper
M L
ML
Melton
D W
DW
D000818:Animals; D002460:Cell Line; D002678:Chimera; D002872:Chromosome Deletion; D005260:Female; D005796:Genes; D005822:Genetic Vectors; D007041:Hypoxanthine Phosphoribosyltransferase / Q000172:deficiency / Q000235:genetics*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D014158:Transcription, Genetic
D007041:Hypoxanthine Phosphoribosyltransferase