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Titolo:
Use of bovine satellite sequences to increase transgene integration by homologous recombination in bovine embryos
Autore:
Rieth, A; Pothier, F; Gagne, M; Sirard, MA;
Indirizzi:
Univ Laval, Dept Anim Sci, Ctr Rech Biol Reprod, St Foy, PQ G1K 7P4, Canada Univ Laval St Foy PQ Canada G1K 7P4 ol Reprod, St Foy, PQ G1K 7P4, Canada
Titolo Testata:
MOLECULAR REPRODUCTION AND DEVELOPMENT
fascicolo: 1, volume: 53, anno: 1999,
pagine: 1 - 7
SICI:
1040-452X(199905)53:1<1:UOBSST>2.0.ZU;2-0
Fonte:
ISI
Lingua:
ENG
Soggetto:
MAMMALIAN-CELLS; DNA; FERTILIZATION; CHROMOSOME; OOCYTES; GENOME; EGGS;
Keywords:
homologous recombination; bovine satellite; transgene integration;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
26
Recensione:
Indirizzi per estratti:
Indirizzo: Sirard, MA Univ Laval, Dept Anim Sci, Ctr Rech Biol Reprod, St Foy, PQ G1K7P4, Canada Univ Laval St Foy PQ Canada G1K 7P4 St Foy, PQ G1K 7P4, Canada
Citazione:
A. Rieth et al., "Use of bovine satellite sequences to increase transgene integration by homologous recombination in bovine embryos", MOL REPROD, 53(1), 1999, pp. 1-7

Abstract

Homologous recombination (HR) has proven to be functional in mammalian embryos. The efficiency of the HR process was tested in bovine zygotes in an attempt to increase the frequency of transgene integration using different lengths of a bovine satellite (BS) DNA flanking both ends of a neo gene marker (called BS500, BS250, and BS50) and neo alone as a control. Pronuclear microinjection at 16-19 hr post insemination (hpi) of the BS500, BS250, BS50or neo fragments at a concentration of 1 ng/mu l resulted in an increasingly negative effect on embryo development. Therefore all microinjections were performed at a single molecular concentration (320 x 10(6) molecules/mu l). After microinjection, the embryos were allowed to develop for 6 days followed by morphological and PCR analysis. The HR event was detected by PCR in 13 of the 26 embryos (43%) that developed beyond the 12-cell stage, 7/22 (31%), 9/27 (33%), and 0/25 (0%) with the BS500, BS250, BS50, and neo constructs respectively. The length of BS homology had no effect on transgene integration. However, embryos injected with BS neo constructs had significantly lower development rates than neo injected zygotes (17% move than 16 cells for BS500; 14% for BS250; 16% for BS50 compared to 32% for neo, P < 0.05,6 replicates). These results demonstrate that BS sequences have a negativeeffect on embryo development and survival regardless of the amount of DNA injected. The use of HR with highly repetitive genomic sequences is therefore a feasible procedure to produce transgenic bovine embryos. (C) 1999 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/11/20 alle ore 08:26:37