Qi Lab CRISPR Plasmids Available from Addgene
A system to regulate a specific gene's expression based on the Type II bacterial CRISPR system for use in either human cells or bacterial cells that is called CRISPR Interference (CRISPRi). For use in human cells a human codon-optimized but enzymatically dead Cas9 nuclease can be targeted to a DNA sequence using a chimeric gRNA. The inactive Cas9/gRNA complex bound to DNA has been shown to interfere with transcriptional elongation, RNA polymerase binding and transcription factor binding. A BFP-Cas9 (enzymatically inactive) fusion construct was also created to help determine successful transformants. For use in bacteria a catalytically dead Cas9 expression plasmid and separate gRNA expression plasmid were also created. In addition, a wild type bacterial Cas9 allows for genome editing in bacterial cells.
These plasmids are described in:
Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression.Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA. Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.. PUBMED.
Via Amir Taheri Ghahfarokhi