Expression-independent gene trap vectors for random and targeted mutagenesis in embryonic stem cells.

ABSTRACT: Promoterless gene trap vectors have been widely used for high-efficiency gene targeting and random mutagenesis in embryonic stem (ES) cells. Unfortunately, such vectors are only effective for genes expressed in ES cells and this has prompted the development of expression-independent vectors. These polyadenylation (poly A) trap vectors employ a splice donor to capture an endogenous gene's polyadenylation sequence and provide transcript stability. However, the spectrum of mutations generated by these vectors appears largely restricted to the last intron of target loci due to nonsense-mediated mRNA decay (NMD) making them unsuitable for gene targeting applications. Here, we present novel poly A trap vectors that overcome the effect of NMD and also employ RNA instability sequences to improve s

SUBMITTER: Tsakiridis A 

PROVIDER: S-EPMC2770648 | biostudies-literature | 2009 Oct

REPOSITORIES: biostudies-literature