Genome Editing with CRISPR/Cas

CRISPR-Cas-based techniques enable precise gene editing workflows, enrichment approaches and in vitro detection assays with easy retargeting. A Cas ribonucleoprotein (RNP) complex activates when its guide RNA binds to a target sequence and its Cas-interacting domain binds to a specific protospacer-adjacent motif (PAM) in a nucleic acid substrate. Delivery formats may incorporate plasmid constructs, viral vectors, mRNA molecules, sgRNA molecules and Cas proteins.

A toolbox to optimize CRISPR-Cas workflows

NEB offers highly characterized Cas nucleases and unique solutions for streamlined RNA synthesis, cloning, and validation of CRISPR-based genome editing. Each Cas endonuclease is characterized for use in common CRISPR applications. For example, EnGen^® Spy Cas9 HF1 (NEB #M0667) is recommended for reduced off-targeting in vivo, EnGen SpRY Cas9 (NEB #M0669) is recommended to expand editable regions in vivo and abolish PAM requirements in vitro and EnGen Lba Cas12a (Cpf1) (NEB #M0653) opens up editing in additional genomic regions and ectothermic organisms. NEBuilder^® HiFi DNA Assembly provides exceptional cloning efficiency to streamline the construction of Cas-sgRNA expression vectors, single sgRNA templates, or sgRNA libraries. Our extensive RNA Synthesis and Modification portfolio supports the synthesis of sgRNA and Cas mRNA. Analysis of genome editing results can progress to validation using our solutions for simple enzymatic detection assays, Sanger sequencing-based detection and next-generation sequencing with NEBNext^® Ultra library prep kits.