Wildtype/Reference Sequence

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Paste a single wildtype/reference sequence (a FASTA header is optional), or upload a single-sequence file (.txt, .fasta).

Edited/Desired Sequence

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Paste a single edited/desired sequence with your intended mutation (a FASTA header is optional), or upload a single-sequence file (.txt, .fasta).

Design Limits:

One sequence per run.
Minimum sequence length: 30 bp total (at least 20 bp upstream, 10 bp downstream of edit).
Prediction algorithm requires at least 99 bp upstream and downstream of the edit.
Maximum sequence length: 2,000 bp (to ensure fast processing).
Maximum file upload size: 10 KB.

PAM sequence

NGG NG

Cut distance to PAM

Spacer length: 20

Spacer GC content (%): 0-100

Prime editing window (the default values are recommended): 1-15

PBS length: 7-16

PBS GC content (%): 0-100

Recommended Tm of PBS sequence (°C): 30

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

Tm-directed PBS length model

Exclude first C in RT template

Dual-pegRNA model

PE3/PE3b nicking sgRNA

Primer Type

pOsU3 pTaU3 pTaU6 pH-nCas9-PPE-V2 Custom

Running Prediciton Model