Guide RNAs with embedded barcodes boost CRISPR-pooled screens.
Genome Biol. 2019 01 24;20(1):20
Authors: Zhu S, Cao Z, Liu Z, He Y, Wang Y, Yuan P, Li W, Tian F, Bao Y, Wei W
We report a new method using re-designed guide RNAs with internal barcodes (iBARs) embedded in their loop regions. Our iBAR approach outperforms the conventional method by producing screening results with much lower false-positive and false-negative rates especially with a high multiplicity of infection (MOI). Importantly, the iBAR approach reduces the starting cells at high MOI significantly with greatly improved efficiency and accuracy compared with the canonical CRISPR screens at a low MOI. This new system is particularly useful when the source of cells is limited or when it is difficult to control viral infection for in vivo screening.
PMID: 30678704 [PubMed – indexed for MEDLINE]