Cell News—Scaling up DNA editing

GFP used to visualize neurons. Image by Jason Snyder.

GFP used to visualize neurons. Image by Jason Snyder.

Easily editing a cell’s native DNA to make specific proteins glow is a cell biologist’s dream. The glowing GFP-tagged proteins can be followed and monitored as they go about their business. But with over 20,000 proteins encoded in the human genome, it’s a daunting task to tag each one. However, Manuel Leonetti and colleagues in ASCB member Bo Huang’s lab at the University of California, San Francisco, found a way to scale up the process without molecular cloning. Using a CRISPR/Cas9 RNA derivative and a split-GFP system they were able to quickly and accurately tag 48 proteins. Plus the tagged proteins were easily isolated for biochemical and proteomic analysis. They believe this method can be used for large-scale protein tagging in human cell lines. Published in PNAS.

About the Author: