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
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