Category: Cellular Technologies

1144-E - A Rapid, Simple, High-throughput Compatible Approach to Generating CRISPR/Cas9 Knock-out Cell Lines

Wednesday, February 7, 2018
11:30 AM - 12:30 PM

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and its associated endonuclease Cas9 have emerged as a revolutionary genome engineering tool.  Using CRISPR/Cas9 to induce mutations in protein encoding genomic DNA regions is a powerful method for studying protein function in the native cellular context.  However, the existing workflows for generating CRISPR/Cas9 knock-out cell lines are cumbersome and labor intensive, comprising of arduous rounds of cloning, limiting dilutions, and surveyor assays performed on dozens if not hundreds of clones. In order for large scale CRISPR/Cas9 studies to become practical and accurate for the genomics community, a streamlined process must be developed and validated at both genomic and protein levels.  Here, we report a novel workflow that leverages the power of droplet digital PCR (ddPCR) and high-resolution melt analysis (HRM) to produce DNA sequence and western blot validated CRISPR/Cas9 knock-out cell lines in under 5 weeks.   

Meiye Wu

Senior Scientist
Bio-Rad Laboratories
Hercules, CA

Meiye Wu, PhD began her involvement with SLAS as an Innovation Award finalist in 2014. Dr. Wu holds a PhD in Biochemistry, MA in Pharmacology, and BA in Neuroscience. She spent nearly a decade at Sandia National Laboratory's Biotechnology and Bioengineering department, spearheading projects to develop automated microfluidics systems and multiplexed assays. Currently, Dr. Wu is a member of the Reagent R&D team at Bio-Rad, Inc, working to integrate traditional enzyme engineering with reagent and platform development for the next generation of genome analysis systems.