High-throughput knock-in coupling gene targeting with the HPRT minigene and Cre-mediated recombination

Tae Moon Kim, Yong Jun Choi, Jun Ho Ko, Paul Hasty

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Single nucleotide polymorphisms (SNPs) may influence protein function possibly contributing to phenotype; yet, for most SNPs their potential influence is unknown. Here, we present a technique in mouse embryonic stem cells that enables high-throughput knockin (the placement of coding sequences adjacent to a specific endogenous promoter). Our methodology utilizes gene targeting with a combination of two selection cassettes (SAβgeo and the HPRT minigene) along with site-specific recombinases (Cre/loxP and FLP/FRT) to efficiently introduce multiple DNA sequences, including enhanced green fluorescent protein (eGFP), adjacent to the DNA topoisomerase 3β (Top3β) promoter. This technology enables rapid and efficient introduction of DNA sequences to a specific location and advances high-throughput analysis of many SNPs with control for expression and genetic background.

Original languageEnglish (US)
Pages (from-to)732-737
Number of pages6
JournalGenesis
Volume46
Issue number12
DOIs
StatePublished - 2008

Keywords

  • Gene targeting
  • Knock-in
  • SNO
  • Top3β
  • eGFP

ASJC Scopus subject areas

  • Genetics
  • Endocrinology
  • Cell Biology

Fingerprint Dive into the research topics of 'High-throughput knock-in coupling gene targeting with the HPRT minigene and Cre-mediated recombination'. Together they form a unique fingerprint.

  • Cite this