Effectiveness of CRISPR-Cas9 Using Pools of Synthetic cRNAs in Screening Experiments
Application Note Feb 09, 2021
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