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 journalArticlepeer-review


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
Issue number1-4
StatePublished - 1993
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


Dive into the research topics of 'Targeting of the gi2α gene in es cells with replacement and insertion vectors'. Together they form a unique fingerprint.

Cite this