Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs
Type
Blog Post
Published
June 2, 2015, 6:06 p.m.
...biological pathways; to observe a phenotype, the modification of multiple genes is often necessary.
Guide ...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end.
PTG construct schematics. Scissors indicate cleavage sites.
After confirming gRNA cleavage...)
How many loci can Cas9 modify?
To test the application of this system to gRNA multiplexing, Xie et al...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct...multiplexing further illustrates the potential of PTGs.
Applications of PTGs
The major advantage of the PTG system... Mar 2. PubMed.
Find the plasmids from this publication at Addgene
Other Techniques for gRNA Multiplexing...