CCTop: an intuitive, flexible and reliable CRISPR/Cas9 target prediction tool
M Stemmer, T Thumberger, M del Sol Keyer… - PloS one, 2015 - journals.plos.org
PloS one, 2015•journals.plos.org
Engineering of the CRISPR/Cas9 system has opened a plethora of new opportunities for site-
directed mutagenesis and targeted genome modification. Fundamental to this is a stretch of
twenty nucleotides at the 5'end of a guide RNA that provides specificity to the bound Cas9
endonuclease. Since a sequence of twenty nucleotides can occur multiple times in a given
genome and some mismatches seem to be accepted by the CRISPR/Cas9 complex, an
efficient and reliable in silico selection and evaluation of the targeting site is key prerequisite …
directed mutagenesis and targeted genome modification. Fundamental to this is a stretch of
twenty nucleotides at the 5'end of a guide RNA that provides specificity to the bound Cas9
endonuclease. Since a sequence of twenty nucleotides can occur multiple times in a given
genome and some mismatches seem to be accepted by the CRISPR/Cas9 complex, an
efficient and reliable in silico selection and evaluation of the targeting site is key prerequisite …
Engineering of the CRISPR/Cas9 system has opened a plethora of new opportunities for site-directed mutagenesis and targeted genome modification. Fundamental to this is a stretch of twenty nucleotides at the 5’ end of a guide RNA that provides specificity to the bound Cas9 endonuclease. Since a sequence of twenty nucleotides can occur multiple times in a given genome and some mismatches seem to be accepted by the CRISPR/Cas9 complex, an efficient and reliable in silico selection and evaluation of the targeting site is key prerequisite for the experimental success. Here we present the CRISPR/Cas9 target online predictor (CCTop, http://crispr.cos.uni-heidelberg.de) to overcome limitations of already available tools. CCTop provides an intuitive user interface with reasonable default parameters that can easily be tuned by the user. From a given query sequence, CCTop identifies and ranks all candidate sgRNA target sites according to their off-target quality and displays full documentation. CCTop was experimentally validated for gene inactivation, non-homologous end-joining as well as homology directed repair. Thus, CCTop provides the bench biologist with a tool for the rapid and efficient identification of high quality target sites.
