Table 3.

List of existing free online tools and open-source local software for sgRNA design.

Name	Graphical user interface	Searches whole genome for targets	Input	Output	Predicts gRNA activity	Ranked list	References
CHOPCHOP	Yes	Yes	Gene ID, chromosome position; also allows gene input	Candidate guide sequences and off-target loci	No	Yes	Montague et al., 2014
CHOPCHOP v2	Yes	Yes (over 200)	Gene ID, chromosome position; also allows gene input	Candidate guide sequences and off-target loci	Yes	Yes	Labun et al., 2016
Cas-OFFFinder	Yes	Yes (over 400)	Guide sequence	Off-target loci for guide sequences	No	No	Bae et al., 2014
CasOT	No (Perl script)	User input	Guide sequence	Off-target loci and additional guide sequences	No	No	Xiao et al., 2014
Benchling	Yes	Yes	DNA sequence or gene name	Candidate guide sequences and off-target loci	No	Yes	Benchling, 2020
CRISPOR	Yes	Yes (over 200 genomes)	DNA sequence	Candidate guide sequences and off-target loci	Yes	Yes	Haeussler et al., 2016