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Intron-Based Single Transcript Unit CRISPR Systems for Plant Genome Editing.

Zhaohui Zhong1, Shishi Liu1, Xiaopei Liu1

  • 1Department of Biotechnology, School of Life Sciences and Technology, Center for Informational Biology, University of Electronic Science and Technology of China, Room 216, Main Building, No. 4, Section 2, North Jianshe Road, Chengdu, 610054, People's Republic of China.

Rice (New York, N.Y.)
|February 5, 2020
PubMed
Summary
This summary is machine-generated.

We developed novel intron-based CRISPR systems for efficient plant genome editing. These single transcript unit (STU) systems enable both singular and multiplexed editing with Cas9 and Cas12a, advancing plant biotechnology.

Keywords:
Cas12aCas9RiceSTU CRISPR 3.0iSTU

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Area of Science:

  • Plant Biotechnology
  • Genome Editing
  • Molecular Biology

Background:

  • Single transcript unit (STU) CRISPR systems offer compact expression of Cas9/Cas12a and guide RNAs.
  • Optimizing STU systems is crucial for routine plant genome editing applications.

Purpose of the Study:

  • To develop and optimize a novel intron-based STU (iSTU) CRISPR system, termed STU CRISPR 3.0.
  • To evaluate the efficacy of different introns and guide RNA processing strategies for plant genome editing.

Main Methods:

  • Evaluated three plant introns for carrying guide RNAs in rice using an eGFP system.
  • Inserted gRNA-containing introns into Cas9 and Cas12a ORFs to test genome editing.
  • Assessed singular and multiplexed genome editing capabilities of iSTU-Cas9 and iSTU-Cas12a systems.

Main Results:

  • Validated proper intron splicing for guide RNA delivery.
  • Demonstrated successful singular and multiplexed genome editing using iSTU-Cas9 and iSTU-Cas12a.
  • Identified effective guide RNA processing strategies for the developed systems.

Conclusions:

  • Successfully developed multiple intron-based STU CRISPR/Cas9 and Cas12a systems for plant genome editing.
  • The developed iSTU systems show promise for advancing plant genome editing technologies.
  • Results provide a foundation for future improvements in STU system design and application.