Protocols for CRISPR Design and Applications

A guide on how to use CRISPR RNP for targeted genome editing, Including the following:

• How to prepare RNA oligos and RNP complex
• How to test RNP efficacy in cells, through either cationic lipid-mediated transfection or electroporation
• How to test RNP efficacy in embryos
• How to test RNP in vitro cutting efficiency

CRISPR Lipofection Protocol for HEK293T/293 cells using Lipofectamine™ 2000 and CRISPRMAX™ Cas9 Transfection Reagents

Cas12a Editing Protocol for HEK 293T/293 Cells Using the Thermo Fisher Neon® Transfection System

Base Editing Protocol for Jurkat cells with Thermo Fisher Scientific Neon™ NxT Electroporation System

Prime Editing Protocol for HEK293T cells with Thermo Fisher Neon® Transfection System

CRISPR Knock-in Protocol for HEK 293T/293 cells with Thermo Fisher Neon^® Transfection System

CRISPR Knock-in Protocol for Jurkat cells with Thermo Fisher Neon^® Transfection System

CRISPR Knock-in Protocol for stimulated Human T Cells with Lonza 4D Nucleofector™ X Unit

CRISPR Knock-in Protocol for Stimulated Human T cells with MaxCyte^® Electroporation Platform

CRISPR Knock-in Protocol for Stimulated Human T-Cells with Thermo Fisher Scientific Neon™ Transfection System

This guideline aims to assist users in designing sgRNA and HDR templates for CRISPR-based gene knockin projects, as well as preparing HDR templates for downstream applications.