Picking the Best CRISPR-Cas9 Targets for Functional Gene Knockout: A Machine Learning Algorithm Based on Both Specificity and Functionality
Poster Sep 24, 2015
Shawn McClelland, Emily M. Anderson, Žaklina Strezoska, Elena Maksimova, Annaleen Vermeulen, Steve Lenger, Tyler Reed, and Anja van Brabant Smith Dharmacon, now part of GE Healthcare, 2650 Crescent Drive, Suite #100, Lafayette, CO 80026, US
Functional gene knockout is an important tool for understanding a gene's role in a system or for specifically manipulating a known system to achieve a desired outcome. Not all gene cleavage events result in functional knockout of the target protein. Here we share important advancements that have helped to achieve the goal of picking the best crRNA targets for functional gene knockout, and not just formation of indels (insertions or the deletions of bases in the DNA).
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE