Home Applications Genome Editing sgRNA Template Construction for Cas9 Gene Editing

sgRNA Template Construction for Cas9 Gene Editing

Return to Genome Editing

Along with Cas9 nuclease, CRISPR experiments require the introduction of an sgRNA containing an approximately 20-base sequence specific to the target DNA 5′ of a non-variable scaffold sequence. sgRNA can be delivered as RNA or by transforming with a plasmid with the sgRNA-coding sequence under a promoter. A number of strategies have been developed to quickly swap out the 20 base sequences allowing convenient sgRNA cloning using NEB products.

sgRNA template construction strategies


Plasmids containing sgRNA sequences can be constructed using a variety of methods. Common to each is the requirement to introduce an approximately 20 base target sequence downstream of a promoter. Guide RNA templates can then be used as templates for in vitro transcription or directly introduced.

Videos

Learn about Genome Editing