CRISPOR is a website that helps select and express CRISPR guide sequences, described in two papers (Gen Biol 2016 and NAR 2018). In its default mode, the user pastes an input DNA sequence and chooses the genome.
CRISPOR is made to be fast and easy on Latch. If anything is confusing, you may refer to Maximilian’s excellent documentation here.
The directory where the files produced by this workflow will be placed. A path can either be selected or if a new path is typed in field Latch will automatically create the folders in the data viewer.
The main output of CRISPOR is a page that shows the annotated input sequence at the top and the list of possible guides in the input sequence at the bottom.
Shown below the input sequence are the guide target sequences, one per PAM. For spCas9, the PAM is NGG and the targets are 20bp long.Column 1 - guide name
this is the position of the PAM on the input sequence and the strand, e.g. “13+”
Column 2 - guide sequence
the sequence of the guide target and the PAM and the link to its “PCR and cloning primers” (see the Primers section
Column 3 - specificity score
a prediction of how much an RNA guide sequence for this target may lead to off-target cleavage somewhere else in the genome.
Column 4 - efficiency scores
the efficiency score is a prediction of how well this target may be cut by its RNA guide sequence.
Column 5 - out-of-frame score
this score (0-100) is a prediction how likely a guide is to lead to out-of-frame deletions.
Column 6 - off-target mismatch counts
the number of possible off-targets in the genome, for each number of mismatches.
Column 7 - off-targets
the locations of all possible off-targets with up to four mismatches, annotated with additional information