Targeting of the gi2α gene in es cells with replacement and insertion vectors

Uwe Rudolph, Philippe Brabet, Jeff Kaplan, Paul Hasty, Allan Bradley, Lutz Birnbaumer

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Five replacement vectors (RV) and one insertion vector (IV) were constructed in which ca. 10 kb of genomic Gi2α sequence, flanked on one (IV) or both (RV) sides by a thymidine kinase (TK) marker, were disrupted by a Neo marker inserted into the Ncol site of exon 3. G418RFIAUR clones corresponding to ca. 4×108 ES cells electroporated with replacement vectors were analyzed and revealed no targeting event. The insertion vector, however, was integrated by a single reciprocal recombination resulting in a duplication of homology (Hit step; G418RFIAUSS, which was lost-together with the plasmid and the TK sequences - by intrachromosomal recombination (Run step; G418RFIAUR). Thus, the Hit and Run strategy can be used with a selectable marker disrupting the targeted gene, giving rise to the same targeted product that would have been expected to arise from a double crossover with a replacement vector.

Original languageEnglish (US)
Pages (from-to)619-637
Number of pages19
JournalJournal of Receptors and Signal Transduction
Volume13
Issue number1-4
DOIs
StatePublished - Jan 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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