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.
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