Rapid Assembly of gRNA Arrays via Modular Cloning in Yeast
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 …
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 …