Guide RNAs with embedded barcodes boost CRISPR-pooled screens
Zhu, Shiyou | Cao, Zhongzheng | Liu, Zhiheng | He, Yuan | Wang, Yinan | Yuan, Pengfei | Li, Wei | Tian, Feng | Bao, Ying | Wei, Wensheng
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.
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