CRISPR genome editing enables precise, intentional, and permanent changes in the DNA of living organisms and cells. This editing technology relies on Cas enzymes and guide RNAs (gRNAs) that are part of the bacterial immune systems found in nature. A gRNA binds the Cas enzyme and directs it to a precise genomic DNA target, where the Cas enzyme then cleaves the DNA. Mutations, gene knockouts, and knock-ins can be precisely placed at the targeted site.
Scientists select an appropriate Cas enzyme and design a gRNA targeting a genomic site. After synthesis, gRNA is combined with the Cas enzyme, forming a ribonucleoprotein (RNP). The RNP is delivered to cells by various means such as electroporation, and genome editing takes place inside cells. For a quantitative assessment, DNA from edited cells can be sequenced by next generation sequencing (NGS).
Efficient CRISPR reagents based on the commonly used Streptococcus pyogenes Cas9 system for lipofection or electroporation experiments. Protospacer adjacent motif (PAM) = NGG.
For additional target sites or for targeting AT-rich regions, use the Acidaminococcus sp. BV3LC CRISPR-Cas12a system in electroporation experiments. Protospacer adjacent motif (PAM) = TTTV. The Alt-R Cas12a (Cpf1) Ultra also can recognize many TTTT PAM sites.
Select from our predesigned gRNAs targeting human, mouse, rat, zebrafish, or nematode genes. For other species, use our proprietary algorithms to design custom gRNAs. For protospacer designs of your own or from publications, use our design checker tool to assess targeting potential before ordering gRNAs that are synthesized using Alt-R gRNA modifications.
Provide basic information about your target site, then use the HDR tool to design and visualize your desired edit within the sequence. The HDR Design Tool will provide the recommended gRNA(s) and HDR donor template for your specifications.