Fecha/hora: 01/06/2024 23:16:02

Biblioteca(s): | INIA La Estanzuela.
Tipo de la producción científica: | Capítulo en Libro Técnico-Científico
Fecha corriente: | 20/07/2022
Fecha de la última actualización: | 20/07/2022
Autoria: | CRISPO, M.; CHENOUARD, V.; DOS SANTOS-NETO P; TESSON, L.; SOUZA-NEVES, M.; HESLAN, J.M.; CUADRO, F.; ANEGÓN, I.; MENCHACA, A.
Afiliación: | MARTINA CRISPO, Laboratory Animal Biotechnology Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay.; VANESSA CHENOUARD, INSERM Centre de Recherche en Transplantation et Immunologie UMR 1064, Transgenesis Rat ImmunoPhenomic Facility (TRIP), Nantes, France.; PEDRO DOS SANTOS-NETO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay.; LAURENT TESSON, INSERM Centre de Recherche en Transplantation et Immunologie UMR 1064, Transgenesis Rat ImmunoPhenomic Facility (TRIP), Nantes, France.; MARCELA SOUZA-NEVES, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay.; JEAN-MARIE HESLAN, INSERM Centre de Recherche en Transplantation et Immunologie UMR 1064, Transgenesis Rat ImmunoPhenomic Facility (TRIP), Nantes, France.; FEDERICO CUADRO, Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay.; IGNACIO ANEGÓN, INSERM Centre de Recherche en Transplantation et Immunologie UMR 1064, Transgenesis Rat ImmunoPhenomic Facility (TRIP), Nantes, France.; JOSE ALEJO MENCHACA BARBEITO, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay./Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay.
Título: | Generation of a Human Deafness Sheep Model Using the CRISPR/Cas System.
Complemento del título: | Chapter 12.
Año de publicación: | 2022
Fuente/Imprenta: | Methods in Molecular Biology, 2022, Volume 2495, Pages 233-258. doi: https://doi.org/10.1007/978-1-0716-2301-5_12
DOI: | 10.1007/978-1-0716-2301-5_12
Idioma: | Inglés

Contenido:

Abstract:
CRISPR/Cas9 system is a promising method for the generation of human disease models by genome editing in non-conventional experimental animals. Medium/large-sized animals like sheep have several advantages to study human diseases and medicine. Here, we present a protocol that describes the generation of an otoferlin edited sheep model via CRISPR-assisted single-stranded oligodinucleotide-mediated Homology-Directed Repair (HDR), through direct cytoplasmic microinjection in in vitro produced zygotes.Otoferlin is a protein expressed in the cochlear inner hair cells, with different mutations at the OTOF gene being the major cause of nonsyndromic recessive auditory neuropathy spectrum disorder in humans. By using this protocol, we reported for the first time an OTOF KI model in sheep with 17.8% edited lambs showing indel mutations, and 61.5% of them bearing knock-in mutations by HDR . The reported method establishes the bases to produce a deafness model to test novel therapies in human disorders related to OTOF mutations.

Palabras-Clave: | Embryo; Genome editing; In vitro fertilization; OTOF; SsODNs.
Thesagro: | OVEJA.
Categoría del tema: | --
Registro original: | INIA La Estanzuela (LE)
Ejemplares: | LE - (103601-1) - PXI - PL - DD - Methods in Molecular Biology/2022 -  -  - DIS