A CRISPR Resource for Individual, Combinatorial, or Multiplexed Gene Knockout
Elsevier (Cell Press)
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Erard, N., Knott, S., & Hannon, G. (2017). A CRISPR Resource for Individual, Combinatorial, or Multiplexed Gene Knockout. Molecular Cell, 67 (2), 348-354.e3. https://doi.org/10.1016/j.molcel.2017.06.030
We have combined a machine-learning approach with other strategies to optimize knockout efficiency with the CRISPR/Cas9 system. In addition, we have developed a multiplexed sgRNA expression strategy that promotes the functional ablation of single genes and allows for combinatorial targeting. These strategies have been combined to design and construct a genome-wide, sequence-verified, arrayed CRISPR library. This resource allows single-target or combinatorial genetic screens to be carried out at scale in a multiplexed or arrayed format. By conducting parallel loss-of-function screens, we compare our approach to existing sgRNA design and expression strategies.
CRISPR, sgRNA, algorithm, combinatorial, screen, expression, Cas9, prediction, lentivirus, library
This work was supported in part by funding from CRUK.
Cancer Research UK (C14303/A17197)
Pew Charitable Trusts (28354)
Cancer Research UK (C14303_do not transfer)
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External DOI: https://doi.org/10.1016/j.molcel.2017.06.030
This record's URL: https://www.repository.cam.ac.uk/handle/1810/265269
Attribution 4.0 International, Attribution 4.0 International, Attribution 4.0 International, Attribution 4.0 International