Thomas Vallim: New Protocol Improves Measurement of AAV-CRISPR Gene Editing
Thomas Vallim, Professor at UCLA, shared a post on LinkedIn about a recent article by Kelsey E. Jarrett et al, published in Star Protocols, adding:
“Turns out your CRISPR editing efficiency might be a lot higher than your Sanger traces are telling you.
In our new STAR Protocols paper, Kelsey Jarrett and the team lay out why standard methods for quantifying AAV-CRISPR editing in the liver undercount things; they’re blind to large DNA structural variants (like whole AAV genomes integrating at the cut site), which can make up nearly half of all editing events.
We worked out a straightforward qPCR trick to capture those alongside conventional INDEL analysis, so labs using AAV-CRISPR in vivo can get a real number instead of a floor.
Also worth noting: 8 of the co-authors on this paper were UCLA undergraduates at the time, providing a great example of how undergrad researchers can contribute to a project.
Proud of this one — nice complement to our Cell Metabolism paper on bile acid metabolism that started this whole thread.”
Title: Protocol for quantification of AAV-CRISPR-mediated hepatic gene editing in mice
Authors: Kelsey E. Jarrett, Rodrigo Baltazar-Nunez, Chloe Anne Borja, Precious Juvie Calderon, Rochelle W. Lai, Angela Cheng, Manasvi Paudel, Gabriella E. Rubert, Anthony Suberski, Owen K. Traina, Sunny Y. Cai, Kalyani Srikanth, William R. Lagor, Elizabeth J. Tarling, Thomas Q. de Aguiar Vallim
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