TY - JOUR AB - CRISPR is a versatile technology for genomic editing and regulation, but the expression of multiple gRNAs in S. cerevisiae has thus far been limited. We present here a simple extension to the Yeast MoClo Toolkit, which enables the rapid assembly of gRNA arrays using a minimal set of parts. Using a dual-PCR, Type IIs restriction enzyme Golden Gate assembly approach, at least 12 gRNAs can be assembled and expressed from a single transcriptional unit. We demonstrate that these gRNA arrays can stably regulate gene expression in a synergistic manner via dCas9-mediated repression. This approach expands the number of gRNAs that can be expressed in this model organism and may enable the versatile editing or transcriptional regulation of a greater number of genes in vivo. AU - McCarty,NS AU - Shaw,WM AU - Ellis,T AU - Ledesma-Amaro,R DO - 10.1021/acssynbio.9b00041 EP - 910 PY - 2019/// SN - 2161-5063 SP - 906 TI - Rapid assembly of gRNA arrays via modular cloning in yeast T2 - ACS Synthetic Biology UR - http://dx.doi.org/10.1021/acssynbio.9b00041 UR - https://www.ncbi.nlm.nih.gov/pubmed/30939239 UR - http://hdl.handle.net/10044/1/70031 VL - 8 ER -