Repositorio UFRO · Omeka S

Improved exogenous DNA uptake in bovine spermatozoa and gene expression in embryos using membrane destabilizing agents in ICSI-SMGT

Primer Autor	Felmer, Ricardo
Co-autores	Sanchez-Villalba, Esther#Elena Arias, Maria#Zambrano, Fabiola#Loren, Pia
Título	Improved exogenous DNA uptake in bovine spermatozoa and gene expression in embryos using membrane destabilizing agents in ICSI-SMGT
Editorial	CAMBRIDGE UNIV PRESS
Revista	ZYGOTE
Lenguaje	en
Resumen	Sperm-mediated gene transfer (SMGT) is a simple, fast, and economical biotechnological tool for producing transgenic animals. However, transgene expression with this technique in bovine embryos is still inefficient due to low uptake and binding of exogenous DNA in spermatozoa. The present study evaluated the effects of sperm membrane destabilization on the binding capacity, location and quantity of bound exogenous DNA in cryopreserved bovine spermatozoa using Triton X-100 (TX-100), lysolecithin (LL) and sodium hydroxide (NaOH). Effects of these treatments were also evaluated by intracytoplasmic sperm injection (ICSI)-SMGT. Results showed that all treatments bound exogenous DNAto spermatozoa including the control. Spermatozoa treated with different membrane destabilizing agents bound the exogenous DNA throughout the head and tail of spermatozoa, compared with the control, in which binding occurred mainly in the post-acrosomal region and tail. The amount of exogenous DNA bound to spermatozoa was much higher for the different sperm treatments than the control (P < 0.05), most likely due to the damage induced by these treatments to the plasma and acrosomal membranes. Exogenous gene expression in embryos was also improved by these treatments. These results demonstrated that sperm membrane destabilization could be a novel strategy in bovine SMGT protocols for the generation of transgenic embryos by ICSI.
Tipo de Recurso	Artículo original
Description	This work was supported by FONDECYT No. 11130724, CONICYT of Chile and Directorate of Research DI17-6003, Universidad de La Frontera. Esther Sanchez and Pia Loren were supported by a Doctoral scholarship by CONICYT, Government of Chile. Este trabajo contó con el apoyo del FONDECYT N.° 11130724, CONICYT de Chile y la Dirección de Investigación DI17-6003 de la Universidad de La Frontera. Esther Sánchez y Pia Loren recibieron una beca de doctorado de CONICYT, Gobierno de Chile.
doi	10.1017/S0967199417000727
Formato Recurso	pdf
Palabras Claves	Cattle# Embryos# ICSI# SMGT# Spermatozoa# Transgenic
Ubicación del archivo	http://dx.doi.org/10.1017/S0967199417000727
Categoría OCDE	Cell Biology# Developmental Biology# Reproductive Biology
Materias	Ganado# embriones# ICSI# SMGT# Espermatozoide# transgénico
Disciplinas de la OCDE	Biotecnología agrícola Ciencia Animal y lechería Biología Reproductiva
Id de Web of Science	WOS:000424128200011
Título de la cita (Recomendado-único)	Improved exogenous DNA uptake in bovine spermatozoa and gene expression in embryos using membrane destabilizing agents in ICSI-SMGT
Identificador del recurso (Mandatado-único)	Artículo original
Versión del recurso (Recomendado-único)	version publicada
Editorial	CAMBRIDGE UNIV PRESS
Revista/Libro	ZYGOTE
Categoría WOS	Biología Celular# Biología del Desarrollo# Biología reproductiva
ISSN	0967-1994
Idioma	en
Referencia del Financiador (Mandatado si es aplicable-repetible)	ANID FONDECYT 11130724#UFRO DI17-6003 ANID FONDECYT 11130724 UFRO DI17-6003
Descripción	This work was supported by FONDECYT No. 11130724, CONICYT of Chile and Directorate of Research DI17-6003, Universidad de La Frontera. Esther Sanchez and Pia Loren were supported by a Doctoral scholarship by CONICYT, Government of Chile.
Formato	pdf
Tipo de ruta	suscripción
Access Rights	metadata
Derechos de acceso	metadata
Página de inicio (Recomendado-único)	5413
Página final (Recomendado-único)	5421