Note: so far gbcloning only supports assemblies of polycistronic crRNAs for LbCas12a.

1) Decide the number of crRNAs that you want to assemble in a single polycistron. Gbcloning supports asselmblies of polycistrons with up to 6 crRNAs. Cas12a has endoribonuclease activity for processing its own crRNAs.

2) Select the most suitable option depending on the number of crRNAs you want to assemble here and upload the target sequences. GB accepts Cas12a target sequences ranging from 20 to 23 nts. Target sequences will be assembled in the final construct in the same order you type them here.

3) Order the synthesis of the crRNA-array you will get as result of 2). We recommend ordering it at GenScript as a Gene Synthesis product and clone it in the pUPD2.

4) Assemble the polycistronic crRNA together with the U6-26-LbCas12aDR promoter fusion here.

5) Combine the TU created at 4) with extra single or polycistronic crRNAs and our recommended TUs/modules for gene editing using the binary assembler.