From a single-nucleotide substitution to a fluorescent reporter tag – a CRISPR knock-in creates precise desired mutations in the genomic DNA near the site where sequence-specific hybridization of an sgRNA recruits the Cas9 nuclease. The desired sequence is delivered into the cell as an HDR template (either an ssDNA oligonucleotide or a donor plasmid), which allows for homology-directed repair (HDR) to occur after Cas9 has made a double-stranded break in the genomic DNA.

Every CRISPR HDR template consists of the desired mutation or insert sequence flanked by left and right homology arms that correspond to the sequence on either side of the CRISPR-induced double stranded break. For small mutations of <50 bp, we suggest using ssDNA oligonucleotide templates, while larger insertions should be handled with one of our dsDNA donor plasmids. Choose to use our Custom CRISPR HDR templates to achieve precise genomic knock-in using the homology-directed repair pathway.

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Applications

1. In-frame fusion tag
   • Tag your gene of interest with a short peptide sequence
   • For this application, an ssDNA oligonucleotide repair template is recommended
2. Point mutation
   • Mutate short sequences or residues within your gene of interest
   • For this application, an ssDNA oligonucleotide repair template is recommended
3. Transgene insertion
   • Insert a larger transgene or reporter into a locus of your choice
   • For this application, a dsDNA vector repair template is recommended

CRISPR Genomic Cleavage Detection Kit

Custom CRISPR sgRNA Lentivectors and Viruses

Cas9 Nuclease Protein