John A. Morris*, Jahan A. Rahman*, Xinyi Guo, Neville E. Sanjana
Quick start guide:
Interactively design pegRNAs/sgRNAs
1) Click on the
Prime editor designer
tab above.
2) Enter a gene symbol or ClinVar ID. The tool will design pegRNAs and
PE3/3b sgRNAs for browsing and download.
Design custom ClinVar prime editors
1) Click on the
Custom ClinVar prime editors
tab above.
2) Input all required design parameters (list of genes, PAM-template, PBS-size, and RTT-size) and provide an e-mail address for design results to be sent to.
3) Click
Design prime editors
and wait for confirmation message. A download link will be sent to the provided e-mail once the design algorithm has finished executing.
Download latest ClinVar prime editors
1) Click on the
Weekly ClinVar prime editors
tab under the
Additional resources
drop-down menu.
2) Bulk download prime editors by clicking the link address or by using 'wget' followed by the link-address (right-click+'copy link address').
For users with design requirements not reflected under the 'Prime editing designer' tab, they may design ClinVar prime editors here in accordance with their needs to be e-mailed to them.
NG-PAM design will increase number of targetable variants and number of pegRNAs/sgRNAs designed per variant, but will take longer to design. PAM-less design will recover virtually all variants, but will take the longest to generate.
Increasing PBS and RTT size will increase number of targetable variants and number of pegRNAs/sgRNAs designed per variant, but will take longer to design.
The custom R scripts may be adapted for the design of prime editors for any species as shown in our tutorial (using an example for prime editing design in zebrafish). After generation of pegRNAs and sgRNAs with the R scripts, we evaluate on-target/off-target scores (and intersections with common dbSNP variants, if relevant) using the scripts listed under the
Flashfry analysis
and
intersection analysis
directories respectively.
For those interested in designing other types of CRISPR reagents for stand-alone use or multiplexed approaches in conjunction with prime editing, please see some of our other online tools: