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Multiplex genome engineering: Methodologies and applications.

Shichang Peng¹, Weiyu Xie², Jie Zhu³

¹Key Laboratory of Biomass Chemical Engineering of Ministry of Education & Zhejiang Key Laboratory of Intelligent Manufacturing for Functional Chemicals, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, Zhejiang, China; ZJU-Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou 311215, Zhejiang, China; Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, Zhejiang, China.

|May 22, 2026

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This summary is machine-generated.

Multiplex genome engineering (MGE) enables simultaneous modification of multiple genes, overcoming previous barriers. This review covers MGE methods and their applications in biotechnology and medicine.

Area of Science:

Genetics
Molecular Biology
Biotechnology

Background:

Single-locus genome editing is established, but modifying multiple genomic sites simultaneously (multiplex genome engineering, MGE) presents challenges.
Advancements in DNA synthesis, genome editing tools, and automation have spurred the development of novel MGE methodologies.

Purpose of the Study:

To review current one-step and iterative multiplex genome engineering methodologies.
To highlight emerging applications of MGE in biomanufacturing, agriculture, and therapeutics.
To analyze limitations and future directions for MGE technologies.

Main Methods:

Analysis of one-step and iterative MGE approaches.
Emphasis on recombineering and CRISPR-Cas systems.
Review of applications across various scientific and industrial sectors.

Keywords:

CRISPR MAGE bioproduction cell therapy iteration multiplex genome engineering xenotransplantation

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Main Results:

A variety of MGE techniques have emerged, addressing previous technological barriers.
MGE is being applied in diverse fields, including basic research, biomanufacturing, agriculture, and therapeutics.
Significant progress has been made in enabling complex genetic modifications.

Conclusions:

Multiplex genome engineering is a rapidly advancing field with transformative potential.
Further optimization of MGE technologies is needed to address system-level biological problems.
MGE is poised to accelerate genetic research and its translation into practical applications.