Faculty Research 1990 - 1999

Title

Positive-negative selection gene targeting with the diphtheria toxin A-chain gene in mouse embryonic stem cells.

Document Type

Article

Publication Date

1993

Keywords

Antigens-CD4: ge, Cell-Line, Diphtheria-Toxin: ge, DNA, Embryo: cy, Genetic-Vectors, Mice, Mutagenesis, Peptide-Fragments: ge, Stem-Cells, SUPPORT-U-S-GOVT-P-H-S

JAX Source

Transgenic Res 1993 Jul;2(4):183-90

Grant

CA10815/CA/NCI, CA18470/CA/NCI, CA21124/CA/NCI

Abstract

The diphtheria toxin A-chain gene was used in a positive-negative selection gene targeting vector to alter the CD4 gene which is transcriptionally silent in mouse embryonic stem cells. Expression of the toxin gene was driven by a constitutively active enhancer, yet the targeting construct exhibited only minimal transient toxicity while enriching for targeted clones 9- to 29-fold. Germline transmission of the stem cell-derived genome was obtained. These data suggest the usefulness of this diphtheria toxin A-chain cassette in replacement-type positive-negative selection vectors. Its potential for novel applications, particularly in the enrichment for 'hit-and-run' insertion-type vectors, is discussed.

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