Fig. 4
From: Gene repression via multiplex gRNA strategy in Y. lipolytica
An overview of the Golden-Brick assembly protocol. Multiplex CRISPRi system contains two main parts, one is JLPC/N-n (or JLRC/N-n) plasmid containing gRNA secretion cassette enabling spacers to be ligated into, the other is PMCS-Multi-CRI vector (classified as dCpf1-Multi and dCas9-Multi) containing the gblock of ‘A’ overhang and ‘T’ overhang enabling to assemble various gRNA secretion cassettes. After being released, these gRNA secretion cassettes were assembled with dCpf1-Multi or dCas9-Multi vector in one step. JLPN-n and JLRN-n plasmids were constructed by replacing synthetic hybrid promoter SCR1′-tRNAGly (JLPC-n and JLRC-n gRNA expression promoter region) with SNR52′-tRNAGly