Protocol for non-invasive delivery of CRISPR RNPs via virus-like particles for mouse model generation

TL;DR

This paper introduces a non-invasive method to edit mouse embryos using CRISPR RNPs delivered via virus-like particles, avoiding physical manipulation and specialized equipment.

Contribution

A novel protocol for CRISPR-VLP-based genome editing in mouse embryos that is non-invasive and equipment-free.

Findings

CRISPR-VLPs enable high-efficiency and heritable genome edits in mouse embryos.

The method reduces off-target effects and eliminates the need for specialized expertise.

It is compatible with zygotes and IVF-derived embryos via co-culture.

Abstract

CRISPR-virus-like particle (VLP)-induced targeted mutagenesis (CRISPR-VIM) enables genome editing in mouse embryos through non-invasive delivery of CRISPR ribonucleoproteins (RNPs) via VLPs, eliminating the need for physical manipulation and specialized expertise. We detail protocols for VLP production, titration, and treatment for diverse genome edits. This protocol is compatible with zygotes and in vitro fertilization (IVF)-derived embryos via simple co-culture, facilitating high-efficiency and heritable mutations with minimized off-target effects, independent of specialized equipment and conducive to reduced animal use.

For complete details on the use and execution of this protocol, please refer to Jeong et al.1

•Detailed steps for producing virus-like particles containing CRISPR ribonucleoproteins•Instructions for safe mouse modeling without physical damage on embryos•Guidance on efficient genetically modified mouse modeling for diverse mutations

Detailed steps for producing virus-like particles containing CRISPR ribonucleoproteins

Instructions for safe mouse modeling without physical damage on embryos

Guidance on efficient genetically modified mouse modeling for diverse mutations

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

CRISPR-virus-like particle (VLP)-induced targeted mutagenesis (CRISPR-VIM) enables genome editing in mouse embryos through non-invasive delivery of CRISPR ribonucleoproteins (RNPs) via VLPs, eliminating the need for physical manipulation and specialized expertise. We detail protocols for VLP production, titration, and treatment for diverse genome edits. This protocol is compatible with zygotes and in vitro fertilization (IVF)-derived embryos via simple co-culture, facilitating high-efficiency and heritable mutations with minimized off-target effects, independent of specialized equipment and conducive to reduced animal use.

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