Literature DB >> 32159922

Simple Protocol for Generating and Genotyping Genome-Edited Mice With CRISPR-Cas9 Reagents.

Almudena Fernández1,2, Matías Morín2,3, Diego Muñoz-Santos1,2, Santiago Josa1,2, Andrea Montero1,2, Marcos Rubio-Fernández1,2, Marta Cantero1,2, Julia Fernández1,2, María Jesús Del Hierro1, Marta Castrillo1, Miguel Ángel Moreno-Pelayo2,3, Lluís Montoliu1,2.   

Abstract

The simple protocol described in this article aims to provide all required information, as a comprehensive, easy-to-follow step-by-step method, to ensure the generation of the expected genome-edited mice. Here, we provide protocols for the preparation of CRISPR-Cas9 reagents for microinjection and electroporation into one-cell mouse embryos to create knockout or knock-in mouse models, and for genotyping the resulting offspring with the latest innovative next-generation sequencing methods.
© 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Designing the best RNA guide for your gene disruption/editing strategy Basic Protocol 2: Preparing and validating CRISPR-Cas9 reagents Basic Protocol 3: Preparing and injecting CRISPR-Cas9 compounds into fertilized mouse oocytes Basic Protocol 4: Genotyping genome-edited mice Support Protocol: Genotyping for CRISPR-generated "indel" mutations. © 2020 John Wiley & Sons, Inc.

Entities:  

Keywords:  CRISPR; animal model; cytoplasmic injection and electroporation; genome editing; in vivo gene editing; mouse model; next-generation sequencing

Year:  2020        PMID： 32159922     DOI： 10.1002/cpmo.69

Source DB:  PubMed          Journal:  Curr Protoc Mouse Biol        ISSN： 2161-2617


  2 in total

1.  CRISPR/Cas9-Mediated Allele-Specific Disruption of a Dominant COL6A1 Pathogenic Variant Improves Collagen VI Network in Patient Fibroblasts.

Authors:  Arístides López-Márquez; Matías Morín; Sergio Fernández-Peñalver; Carmen Badosa; Alejandro Hernández-Delgado; Daniel Natera-de Benito; Carlos Ortez; Andrés Nascimento; Daniel Grinberg; Susanna Balcells; Mónica Roldán; Miguel Ángel Moreno-Pelayo; Cecilia Jiménez-Mallebrera
Journal:  Int J Mol Sci       Date:  2022-04-16       Impact factor: 6.208

2.  Screening and validation of genome-edited animals.

Authors:  Rosie K Bunton-Stasyshyn; Gemma F Codner; Lydia Teboul
Journal:  Lab Anim       Date:  2021-07-01       Impact factor: 2.471
  2 in total

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