RRC ID 4768
Author Kaartinen V, Nagy A.
Title Removal of the floxed neo gene from a conditional knockout allele by the adenoviral Cre recombinase in vivo.
Journal Genesis
Abstract Conditional and tissue specific gene targeting using the Cre-loxP recombination system in combination with established ES cell techniques has become a standard for in vivo loss of function studies. In a typical flox and delete gene targeting strategy, the loxP-neo-loxP cassette is inserted into an intron and an additional loxP site is located in one of the homology arms so that loxP sites surround a functionally essential part of the gene. The neo cassette in usually removed by transient expression of the Cre recombinase in ES cells to avoid selection gene interference and genetic ambiquity. However, this causes a significant increase in manipulation of ES cells and often compromises ES cell pluripotency. Here we describe a method in which the floxed neo gene is removed from a knockout allele by infecting 16-cell-stage morulae by the recombinant Cre adenovirus. This virus provides only transient Cre expression and does not integrate into the mouse genome. Produced mosaic mice transmitted the desired allele without the neo cassette with high frequency to their offspring. This method is rapid and easy and does not require any special equipment. Moreover, because superovulated mice can be used as donors, this method does not necessitate a large number of mice.
Volume 31(3)
Pages 126-9
Published 2001-11-1
DOI 10.1002/gene.10015
PII 10.1002/gene.10015
PMID 11747203
MeSH Adenoviridae / genetics* Alleles Animals Crosses, Genetic Female Gene Deletion Gene Targeting / methods Gene Transfer Techniques Genetic Vectors Integrases / metabolism* Kanamycin Kinase / genetics* Mice Mice, Inbred C57BL Mice, Inbred CBA Mice, Knockout Mice, Transgenic Viral Proteins / metabolism*
IF 1.76
Times Cited 47
DNA material AxCANCre (RDB01748)