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Pooled library screening with multiplexed Cpf1 library.

Jintan Liu1,2, Sanjana Srinivasan3,4, Chieh-Yuan Li3,4

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Summary
This summary is machine-generated.

We developed a novel CRISPR screening method using AsCpf1, creating the smallest whole-genome knockout library. This efficient approach minimizes library size while maintaining gene targeting accuracy.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • CRISPR-Cas9 technology enables functional genomics screening.
  • Current CRISPR libraries can be large and complex.
  • AsCpf1 offers inherent multiplexing capabilities.

Purpose of the Study:

  • To develop a multiplexed, high-throughput screening strategy using AsCpf1.
  • To minimize library size without compromising gene targeting efficiency.
  • To create the smallest whole-genome CRISPR knockout library for humans.

Main Methods:

  • Utilized the multiplexing capability of AsCpf1 for library construction.
  • Developed a multiplexed screening strategy.
  • Constructed the Mini-human whole-genome CRISPR knockout library (17,032 constructs).

Main Results:

  • AsCpf1-based multiplexed libraries perform comparably to monocistronic CRISPR/Cas9 libraries.
  • Achieved efficient gene targeting with a minimized library size.
  • The Mini-human library is the smallest whole-genome CRISPR knockout library for humans.

Conclusions:

  • AsCpf1 is suitable for functional genomics screenings.
  • AsCpf1-based multiplexed libraries offer an efficient alternative to conventional CRISPR/Cas9 libraries.
  • The Mini-human library provides a powerful tool for human genome-wide studies.