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 language | English (US) |
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Pages (from-to) | 732-737 |
Number of pages | 6 |
Journal | Genesis |
Volume | 46 |
Issue number | 12 |
DOIs | |
State | Published - 2008 |
Keywords
- Gene targeting
- Knock-in
- SNO
- Top3β
- eGFP
ASJC Scopus subject areas
- Genetics
- Endocrinology
- Cell Biology