Enhanced gene trapping in mouse embryonic stem cells
Gene trapping is used to introduce insertional mutations into genes of mouse embryonic stem cells (ESCs). It is performed with gene trap vectors that simultaneously mutate and report the expression of the endogenous gene at the site of insertion and provide a DNA tag for rapid identification of the disrupted gene. Gene traps have been employed worldwide to assemble libraries of mouse ESC lines harboring mutations in single genes, which can be used to make mutant mice. However, most of the employed gene trap vectors require gene expression for reporting a gene trap event and therefore genes that are poorly expressed may be under-represented in the existing libraries. To address this problem, we have developed a novel class of gene trap vectors that can induce gene expression at insertion sites, thereby bypassing the problem of intrinsic poor expression. We show here that the insertion of the osteopontin enhancer into several conventional gene trap vectors significantly increases the gene trapping efficiency in high-throughput screens and facilitates the recovery of poorly expressed genes.
|Author:||Frank Schnütgen, Jens Hansen, Silke De-Zolt, Carsten Horn, Marcus Lutz, Thomas Floss, Wolfgang Wurst, Patricia Ruiz Noppinger, Harald von Melchner|
|Date of Publication (online):||01.10.2008|
|Year of first Publication:||2008|
|Publishing Institution:||Univ.-Bibliothek Frankfurt am Main|
|Source:||Nucleic Acids Research, 2008, 1–8 ; doi:10.1093/nar/gkn603|
|Dewey Decimal Classification:||570 Biowissenschaften; Biologie|
(c) 2008 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
|Licence (German):||Veröffentlichungsvertrag für Publikationen ohne Print on Demand|