Effectiveness of CRISPR-Cas9 Using Pools of Synthetic cRNAs in Screening Experiments

Gene knockout, using CRISPR-Cas9, has emerged as a powerful technology for loss-of-function screening. Chemical synthesis of guide RNAs for CRISPR-Cas9 gene editing allows for accurate and rapid production of CRISPR libraries and enables screening in an arrayed “one-gene-per-well” format. The pooling of several crRNAs targeting the same gene into a single reagent has been shown to simplify the screening process.

Having several crRNAs to guide the editing of the gene at different positions could result in a more thorough functional gene knockout, leading to a more robust phenotype that would improve hit identification. However, there are several potential concerns for using crRNA pools.

Download this app note to:

• Address concerns and learn about key considerations related to the use of crRNA pools
• Discover an example that shows pooling crRNAs to the same gene target provides robust functional knockout
• Reveal ways to ensure crRNA pools work successfully in phenotypic screening