In vivo site-specific recombination using the beta-rec/six system.
The prokaryotic beta serine recombinase (beta-rec) catalyzes site-specific recombination between two directly oriented six sites (93 bp) in mammalian cells, both in episomal and in chromosomally integrated substrates. The beta-rec/six exclusive intramolecular site-specific recombination (SSR) system has been proposed as a suitable approach when several independently controlled recombination events are needed in a single cell. Here we explored the use of the beta-rec/six system for selective induction of genome-targeted modifications. We generated and analyzed mouse transgenic lines (Tgbeta) expressing beta-rec under the control of the Lck promoter. beta-rec activity was demonstrated, and there was no evidence of alterations to thymic or peripheral T cell development. We developed two transgenic mouse lines harboring different target sequences (Tgrec and KOsix) and analyzed the effect of beta-rec expression on these animals. The results indicate that the beta-rec/six SSR system is functional for in vivo gene-targeting applications.
Servert
Pilar
P
Departamento de Inmunología y Oncología, Campus Universidad Autónoma de Madrid, Madrid, Spain.
Díaz
Vicente
V
Lucas
Daniel
D
de la Cueva
Teresa
T
Rodriguez
Maria
M
García-Castro
Javier
J
Alonso
Juan
J
Martínez-A
Carlos
C
González
Manuel
M
Bernad
Antonio
A
D000818:Animals; D004254:DNA Nucleotidyltransferases / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D009928:Organ Specificity; D011995:Recombination, Genetic; D013950:Thymus Gland / Q000201:enzymology
D004254:DNA Nucleotidyltransferases; C077570:Site-specific recombinase