Positive control crRNAs can be used to show that Cas12a editing is occurring in your experiments, which can be useful when you are optimizing RNP delivery conditions or if you need to troubleshoot your experiments.
Attention: Unlike S. pyogenes Cas9, which cleaves most potential NGG PAM sites to some degree, some of the tested TTTV sites show no cleavage by A.s. Cas12a nuclease. We recommend using positive control crRNAs to establish that your cells can be edited by Cas12a. In addition, we suggest testing 3 or more crRNAs per target gene.
Our scientists have designed and tested positive control crRNAs targeting HPRT (Figure 4). To order, copy and paste the appropriate sequence in the Cas12a crRNA ordering page:
Human HPRT1, Cas12a Positive Control crRNA: GGTTAAAGATGGTTAAATGAT
Mouse Hprt, Cas12a Positive Control crRNA: GGATGTTAAGAGTCCCTATCT
Rat Hprt1, Cas12a Positive Control crRNA: ACCGCCCCCCCCATACCCCAA
Our scientists have also designed and tested PCR primers (Alt-R HPRT PCR Primer Mixes for human, mouse, or rat) for use with the Alt-R Genome Editing Detection Kit to detect editing or estimate editing efficiency in samples transfected with the positive
control HPRT crRNAs.