Generation of a non-leaky heat shock-inducible Cre line for conditional Cre/lox strategies in zebrafish.
Dev Dyn
; 240(1): 108-15, 2011 Jan.
Article
en En
| MEDLINE
| ID: mdl-21117149
ABSTRACT
Cre-mediated site-specific recombination has emerged as an indispensable tool for the precise manipulation of the mammalian genome. Recently, we showed that Cre is also highly efficient in zebrafish and temporal control of recombination can be achieved by using the ligand-inducible CreER(T2). Previous attempts have been made to control recombination by using the temperature inducible hsp70l promoter to conditionally drive the expression of Cre or EGFP-Cre, respectively. However, in this study we demonstrate that the hsp70l promoter possesses a basal leakiness resulting in Cre-mediated recombination even at permissive temperatures. In order to prevent non-conditional recombination, we combined the hsp70l promoter with a mCherry-tagged ligand-inducible CreER(T2). At permissive temperatures and in the absence of the ligand tamoxifen (TAM), no non-conditional recombination is observed indicating tight regulation of CreER(T2). Instead, comprehensive site-specific recombination is mediated following heat induction and administration of TAM.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Pez Cebra
/
Activación Transcripcional
/
Técnicas de Transferencia de Gen
/
Integrasas
/
Vectores Genéticos
/
Calor
Tipo de estudio:
Evaluation_studies
Límite:
Animals
Idioma:
En
Revista:
Dev Dyn
Asunto de la revista:
ANATOMIA
Año:
2011
Tipo del documento:
Article
País de afiliación:
Alemania