Alt-R CRISPR-Cas12a (Cpf1) crRNA and design tips
The Alt-R CRISPR-Cas12a (Cpf1) crRNA is a single, 40–44 base, guide RNA, comprising a 20 base constant region (loop domain) and
a 20–24 base target-specific region (protospacer domain). We typically recommend a 21 base protospacer domain for optimal
activity. All Alt-R CRISPR-Cas12a (Cpf1) crRNAs are synthesized with proprietary chemical modifications, which protect the crRNA
from degradation by cellular RNases and further improve on-target editing performance. For crRNAs used with A.s. Cas12a,
identify locations in your target region with the protospacer adjacent motif (PAM) sequence, TTTV, where V is A, C, or
G. Your Alt-R CRISPR-Cas12a (Cpf1) crRNA will bind to the DNA strand opposite to the PAM sequence (Figure 2). Do not include the
PAM sequence in your crRNA design. An example of a correct crRNA sequence is shown in Figure 3.
After you enter your 20–24 base target sequence, 20 additional bases and the necessary modifications will automatically be
added by the order entry system for a total of 40–44 RNA bases. These additional bases and modifications are necessary
to create a complete Alt-R CRISPR-Cas12a (Cpf1) crRNA. The system will also convert the final sequence to RNA—enter DNA bases
only into the ordering tool (Figure 3).