Premium
A reliable lacZ expression reporter cassette for multipurpose, knockout‐first alleles
Author(s) -
Testa Giuseppe,
Schaft Julia,
van der Hoeven Frank,
Glaser Stefan,
Anastassiadis Konstantinos,
Zhang Youming,
Hermann Thomas,
Stremmel Wolfgang,
Stewart A. Francis
Publication year - 2004
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.20012
Subject(s) - conditional gene knockout , biology , gene knockout , gene targeting , homologous recombination , genetics , gene , gene knockin , allele , selectable marker , cre lox recombination , reporter gene , function (biology) , knockout mouse , transgene , gene expression , genetically modified mouse , phenotype
Alteration of the mouse genome through homologous recombination in embryonic stem (ES) cells is the most accurate and versatile way to dissect gene function in a vertebrate model. Most often, a selectable marker is used to create a knockout allele by replacing an essential part of the gene. However, knockout strategies are limited because the mutation is present constitutively. Conditional approaches based on the Cre‐ loxP site‐specific recombination (SSR) system address this limitation; however, it requires that all parts of the targeted gene remain in ES cells. Here we report success with a “knockout‐first” strategy that ablates gene function by insertion of RNA processing signals without deletion of any of the target gene. Incorporation of site‐specific recombination target sites creates a multipurpose allele for both knockout and conditional applications. genesis 38:151–158, 2004. © 2004 Wiley‐Liss, Inc.