RRC ID |
4768
|
著者 |
Kaartinen V, Nagy A.
|
タイトル |
Removal of the floxed neo gene from a conditional knockout allele by the adenoviral Cre recombinase in vivo.
|
ジャーナル |
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.
|
巻・号 |
31(3)
|
ページ |
126-9
|
公開日 |
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
|
引用数 |
47
|
WOS 分野
|
GENETICS & HEREDITY
DEVELOPMENTAL BIOLOGY
|
リソース情報 |
遺伝子材料 |
AxCANCre (RDB01748) |