Simple protocol for generating and genotyping genome‐edited mice with CRISPR‐Cas9 reagents
A Fernández, M Morín, D Muñoz‐Santos… - Current Protocols in …, 2020 - Wiley Online Library
A Fernández, M Morín, D Muñoz‐Santos, S Josa, A Montero, M Rubio‐Fernández…
Current Protocols in Mouse Biology, 2020•Wiley Online LibraryThe 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 …
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 …
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
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