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Related Concept Videos

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Updated: Nov 15, 2025

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Developing GDi-CRISPR System for Multi-copy Integration in Saccharomyces cerevisiae.

Zi-Xu Zhang1, Yu-Zhou Wang1, Ying-Shuang Xu1

  • 1School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, 2 Xuelin Road, Qixia District, Nanjing, Jiangsu Province, China.

Applied Biochemistry and Biotechnology
|March 4, 2021
PubMed
Summary

A new CRISPR-based system, GDi-CRISPR, enables efficient multi-copy gene integration in Saccharomyces cerevisiae. This low-cost tool achieves high copy numbers rapidly, advancing biofuel and chemical production.

Keywords:
CRISPR/CasDelta site integrationMulti-copySaccharomyces cerevisiae

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Biotechnology

Background:

  • Saccharomyces cerevisiae is crucial for biofuel and chemical production.
  • Stable expression of target products requires multi-gene integration into the S. cerevisiae chromosome.
  • Existing CRISPR multi-copy integration methods are costly and inefficient.

Purpose of the Study:

  • To develop a low-cost, user-friendly tool for multi-copy gene integration in S. cerevisiae.
  • To improve the efficiency and speed of integrating multiple genes into the yeast genome.

Main Methods:

  • Screened 21 Cas proteins for optimal cleavage activity in S. cerevisiae.
  • Investigated donor translocation order to optimize integration copy number.
  • Developed the Gene Drive delta site integration CRISPR (GDi-CRISPR) system by integrating gRNA into donor fragments.

Main Results:

  • Eight Cas proteins demonstrated gene editing capability.
  • Optimized donor translocation achieved 4 copies.
  • The GDi-CRISPR system yielded 6 copies within 5-6 days without high-throughput screening or resistance markers.

Conclusions:

  • The GDi-CRISPR system offers an efficient and cost-effective solution for multi-copy integration in S. cerevisiae.
  • This technology has the potential to significantly advance the development of S. cerevisiae as a cell factory.