CRISPROne

Design of Prime Editing guide RNAs

Prime editing (PE) tools that consist of a reverse transcriptase linked with Cas9 nickase are capable of generating targeted insertions, deletions, and base conversions without producing DNA double strand breaks or requiring any donor DNA. More ....

Advantages:

Relatively less restricted by PAM / Universal and easy / Precise and flexible

Note:

The designed mutations or corrections are marked as "Substitution (a/b), Insertion (+c), Deletion (-c)". "a" is the original sequence and "b" is the designed mutation sequence, "c" is the deletion (-) or insertion (+) sequence.

Input Sequence

PAM Type See notes on enzymes in the help

Target Genome: More Information of Genomes Metadata

Customized PAM (Need to select Customized PAM in PAM Type )

Please input a valid PAM DNA Sequence.

sgRNA module

Cut distance to PAM

Spacer length

Note: For a Customized PAM select Customized PAM: 5'-XXX-3' in PAM Type and then set sgRNA module , Cut distance to PAM and Spacer length .

pegRNA Spacer GC content(%):

PBS length (bp):

PBS GC content (%):

Recommended Tm of PBS sequence (℃):

Homologous RT template length (the default values are recommended) (bp): 7-16:

Exclude first C in RT template

Dual-pegRNA model

ngRNA spacers (same PAM with pegRNA)

Distance of secondary nicking sgRNAs (ngRNA) to pegRNA (bp):

pegLIT

Linker Pattern

Incorporated structured RNA motifs (Read of article or Help in this site.)

pOsU3 pTaU3 pH-nCas9-PPE-V2/pH-ePPE

Forward primer (5'-3'): + Spacer sequence +

Reverse primer (5'-3'): + PBS and RT template sequence +

Design of Prime Editing guide RNAs

Mandatory Parameters

Optional Parameters

Primer Design