*Scoring is available for 20-nt type II sgRNAs. See Doench et al., 2014 for details.
High scores may indicate more potent sgRNAs. However, some potent sgRNAs have scores < 0.1 and a number of successful sgRNAs had scores between 0.1 and 0.2.
Therefore, potential experimental utility and subsequent experimental verification should guide the choice of sgRNAs.
Off-target analysis parameters: GT-Scan: No restrictions on PAM sequence or target length. Adjust the target rule on the submission form.Cas-OFFinder: Length should be 20bp for SpCas9 (PAM = 'NGG'), 18bp for StCas9 (PAM = 'NNAGAAW'), and 24bp for NmCas9 (PAM = 'NNNNGMTT').
When binding a guide RNA, wild-type Cas9 produces a single double-strand cut, whereas D10A Cas9 mutant (nickase) generates single-stranded nicks in DNA and therefore requires two guide RNAs to induce a cut in DNA.Please select the spacing between two gRNA target sites from 0 to 20.
Please enter your sequence(s) in the text box below
*Please enter one or more sequences in FASTA format, each smaller than 50 kb and having unique names.
Search of target sites will be performed independently in each sequence.
OR enter identifiers of genes or transcripts to run the program on.
*Refseq sequences can be from any species
*The program supports Ensembl gene and transcript identifiers, gene symbols as well as Refseq cDNA accession numbers (if available).
CRISPR MultiTargeter was developed by Sergey Prykhozhij at the IWK Health Centre and Dalhousie University. The design of the logo and the buttons was by Vinothkumar Rajan. The latest update was in May 2019.