Haoquan Wu, Ph.D., assistant professor in the TTUHSC at El Paso Center of Excellence in Infectious Diseases recently presented at the Global Technology Community Non-Coding RNAs and RNAi Research & Therapeutics Conference 2014 in San Diego. Dr. Wu was also invited to participate on the scientific advisory board for the meeting.
Here is a synopsis of what Dr. Wu presented: “Currently, genome-wide functional screening in cultured cells is mostly performed using RNAi, in which gene silencing is incomplete. The genome-editing technology based on the type II prokaryotic CRISPR (clustered regularly interspaced palindromic repeats)-Cas9 endonuclease system has potential utility in genome-wide knockout screening. Here, we designed a strategy to perform genome-wide screening in human cell lines based on this system. Using a library containing 77,406 unique single-guide RNA (sgRNA) sequences targeting 20,121 genes, delivered with a lentiviral vector in combination with transient expression of Cas9, we performed screens to identify the genes that are dispensable for cellular survival and yet essential for HIV-induced cell killing. One of the most serious problems in genome-wide functional screening is the high noise level that significantly reduces the data quality. Here we identified major sources of noise and increased screening specificity. The results showed that our approach can be used to perform genome-wide knockout screens with high specificity and sensitivity,” said Dr. Wu.
Learn more about the meeting at https://www.gtcbio.com/conferences/rna-agenda.