Characterization of Cre-mediated cassette exchange after plasmid microinjection in fertilized mouse oocytes.
Genesis
; 27(4): 153-8, 2000 Aug.
Article
em En
| MEDLINE
| ID: mdl-10992325
ABSTRACT
DNA cassette exchange methodologies are powerful tools for rapid modification of defined genomic loci. Until now, most studies have focused on transfected cell lines that were subjected to positive and negative selection to enrich for correctly recombined clones. This is the first report on Cre-mediated cassette exchange (CMCE) in early mouse embryos, a nonselectable biological system. A Cre expression plasmid and a replacement plasmid were introduced into fertilized oocytes by plasmid microinjection and recombination products were investigated. Both, replacement plasmid and oocyte genome carried heterospecific lox sites (lox511 and loxP) to direct CMCE toward integration. Here, we demonstrate the general feasibility of CMCE in early mouse embryos and characterize the system with respect to integration efficiency, competing reactions and parameters affecting recombination rates.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Oócitos
/
Proteínas Virais
/
Mutagênese Insercional
/
Integrases
/
Microinjeções
Limite:
Animals
Idioma:
En
Revista:
Genesis
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
Alemanha
País de publicação:
EEUU
/
ESTADOS UNIDOS
/
ESTADOS UNIDOS DA AMERICA
/
EUA
/
UNITED STATES
/
UNITED STATES OF AMERICA
/
US
/
USA