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).
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Genome-wide association studies (GWAS) have identified more than 100 genetic loci associated with type 2 diabetes. The majority of these are located in the intergenic or intragenic regions suggesting that the implicated variants may alter chromatin conformation. This, in turn, is likely to influence the expression of nearby or more remotely located genes to alter beta cell function. At present, however, detailed molecular and functional analyses are still lacking for most of these variants. We recently analysed one of these loci and mapped five causal variants in an islet-specific enhancer cluster within the STARD10 gene locus. Here, we aimed to understand how these causal variants influence b-cell function by alteration of the chromatin structure of enhancer clusterREAD MORE
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