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Guide RNAs with embedded barcodes boost CRISPR-pooled screens.

ABSTRACT: 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.

SUBMITTER: Zhu S

PROVIDER: S-EPMC6345036 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Guide RNAs with embedded barcodes boost CRISPR-pooled screens.

Zhu Shiyou S Cao Zhongzheng Z Liu Zhiheng Z He Yuan Y Wang Yinan Y Yuan Pengfei P Li Wei W Tian Feng F Bao Ying Y Wei Wensheng W

Genome biology 20190124 1

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. T ...[more]

PMID: 30678704

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