| First Author | Kim TM | Year | 2008 |
| Journal | Genesis | Volume | 46 |
| Issue | 12 | Pages | 732-7 |
| PubMed ID | 18932256 | Mgi Jnum | J:148743 |
| Mgi Id | MGI:3846297 | Doi | 10.1002/dvg.20439 |
| Citation | Kim TM, et al. (2008) High-throughput knock-in coupling gene targeting with the HPRT minigene and Cre-mediated recombination. Genesis 46(12):732-7 |
| abstractText | 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 knock-in (the placement of coding sequences adjacent to a specific endogenous promoter). Our methodology utilizes gene targeting with a combination of two selection cassettes (SAbetageo 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 3beta (Top3beta) 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. |
