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Related Experiment Video

Updated: Dec 23, 2025

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit
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A modular cloning toolkit for genome editing in plants.

Florian Hahn1, Andrey Korolev1,2, Laura Sanjurjo Loures1

  • 1Plant Sciences Department, Rothamsted Research, Harpenden, AL5 2JQ, UK.

BMC Plant Biology
|April 25, 2020
PubMed
Summary

This study introduces an expanded CRISPR/Cas cloning toolkit featuring 103 modules for plant genome editing. The toolkit simplifies the assembly of CRISPR/Cas components and guide RNAs, enhancing modular cloning efficiency.

Keywords:
CRISPRCas9Genome editingGolden GateMoCloPlant

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Area of Science:

  • Plant biotechnology
  • Molecular biology
  • Genome editing

Background:

  • CRISPR/Cas is a key genome editing tool in plants.
  • Efficient delivery of multiple expression units necessitates streamlined cloning methods.
  • Modular cloning (MoClo) and Golden Gate (GG) facilitate standardized genetic part assembly.

Purpose of the Study:

  • To present an expanded cloning toolkit for CRISPR/Cas applications in plants.
  • To provide standardized, interchangeable genetic parts for constructing expression units and multigene constructs.

Main Methods:

  • Development of a comprehensive toolkit with 103 modules.
  • Inclusion of promoters for CRISPR/Cas expression in monocots and dicots.
  • Design of modules for rapid assembly of tRNA-sgRNA polycistronic units.

Main Results:

  • The toolkit contains CRISPR/Cas nucleases and guide RNA backbones.
  • Promoters enable CRISPR/Cas and guide RNA expression in diverse plant types.
  • A novel system allows PCR-free assembly of tRNA-sgRNA units, functional in wheat protoplasts.

Conclusions:

  • The CRISPR/Cas toolkit is a valuable resource for plant science research.
  • It promotes wider adoption of CRISPR/Cas genome editing and modular cloning.
  • Facilitates advanced genetic engineering in plants.