Vesicles cytoplasmic injection: an efficient technique to produce porcine trandgene expressing embryos

LUCHETTI, CG.; BEVACQUA, RJ.; LORENZO, MS.; TELLO, MF.; WILLIS,M.; BUEMO, CP.; LOMBARDO, DM.; SALAMONE, D.

REPRODUCTION IN DOMESTIC ANIMALS (1990)

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2016 vol. 51 p. 501 - 508

0936-6768

The use of vesicles co-incubated with plasmids showed to improve the efficiency of cytoplasmic injection of transgenes in cattle. Here this technique was tested as a simplified alternative for transgenes delivery in porcine zygotes. To this aim, cytoplasmic injection of the plasmid alone was compared to the injection with plasmids co-incubated with vesicles both in diploid parthenogenic and IVF zygotes. The plasmid pcx-egfp was injected circular (CP) at 3, 30 and 300ng/mL and linear (LP) at 30ng/μL. The experimental groups using parthenogenetic zygotes were: CP naked at 3 ng/mL (N=105), 30ng/mL (N=95) and 300ng/mL (N=65); Sham (N=105); control not injected (N=223); LP naked at 30ng/mL (N=78); LP vesicles (N=115) and Sham vesicles (N=59). For IVF zygotes: LP naked (N=44) LP vesicles (N=94), sham (N=59) and control (N=79). Cleavage, blastocyst and GFP+ rates were analyzed by Fisher?s test (P