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Related Experiment Video

Updated: Feb 24, 2026

A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
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Dense sgRNA Library Construction Using a Molecular Chipper Approach.

Jijun Cheng1,2, Wen Pan1,2, Jun Lu1,2,3,4

  • 1Department of Genetics, Yale University School of Medicine, New Haven, Connecticut, USA.

Bio-Protocol
|August 25, 2017
PubMed
Summary
This summary is machine-generated.

We developed a Molecular Chipper protocol to create dense single-guide-RNA (sgRNA) libraries for CRISPR genetic screens. This method efficiently targets noncoding genomic regions, enabling functional element discovery.

Keywords:
CRISPR-Cas9Molecular chipperNon-coding genomeReporter screensgRNA library

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Area of Science:

  • Genomics
  • Molecular Biology
  • CRISPR Technology

Background:

  • CRISPR genetic screens are vital for identifying genetic regulators in coding and noncoding genomic regions.
  • Interrogating noncoding elements necessitates dense, cost-effective, and customizable sgRNA libraries.

Purpose of the Study:

  • To present the Molecular Chipper protocol for generating dense sgRNA libraries.
  • To enable efficient interrogation of functional elements within specific genomic regions.

Main Methods:

  • The Molecular Chipper protocol uses random DNA fragmentation.
  • A Type III restriction enzyme is employed to generate sgRNA libraries.
  • This method allows for dense coverage of selected genomic regions.

Main Results:

  • The protocol successfully generates dense sgRNA libraries from input DNA.
  • It provides a flexible and efficient method for targeting noncoding regions.

Conclusions:

  • The Molecular Chipper protocol is a valuable tool for creating custom sgRNA libraries.
  • This facilitates comprehensive functional genomic screens, particularly for noncoding elements.