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Related Experiment Video

Updated: Feb 26, 2026

A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
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A CRISPR Resource for Individual, Combinatorial, or Multiplexed Gene Knockout.

Nicolas Erard1, Simon R V Knott2, Gregory J Hannon3

  • 1Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.

Molecular Cell
|July 22, 2017
PubMed
Summary
This summary is machine-generated.

We developed a genome-wide CRISPR library using machine learning and a novel multiplexed sgRNA strategy to improve gene knockout efficiency for large-scale genetic screens.

Keywords:
CRISPRCas9algorithmcombinatorialexpressionlentiviruslibrarypredictionscreensgRNA

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • The CRISPR/Cas9 system is a powerful tool for gene editing.
  • Optimizing knockout efficiency and enabling multiplexed genetic screens are crucial for large-scale functional genomics.

Purpose of the Study:

  • To develop an optimized CRISPR library for efficient gene knockout.
  • To enable scalable, high-throughput genetic screens using a novel sgRNA strategy.

Main Methods:

  • Combined machine learning with CRISPR/Cas9 for optimizing knockout efficiency.
  • Developed a multiplexed sgRNA expression strategy for single and combinatorial gene targeting.
  • Constructed a genome-wide, sequence-verified, arrayed CRISPR library.

Main Results:

  • The developed CRISPR library facilitates efficient gene ablation.
  • The multiplexed sgRNA strategy allows for combinatorial targeting.
  • The library supports both single-target and combinatorial genetic screens at scale.

Conclusions:

  • The new CRISPR library and sgRNA strategy enhance the efficiency and scalability of loss-of-function genetic screens.
  • This resource provides a valuable tool for comprehensive genomic studies.