Related Experiment Video
Updated: May 24, 2026
A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
Published on: July 12, 2017
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.
Abstract:
To facilitate the study of applied genetics and enable a rapid translation of genetic insights, it is highly desirable to concurrently modify many genomic loci in an organism of interest. While single-locus editing is well-established and technically straightforward, multiplex genome engineering (MGE) poses significant technological barriers. With the convergence of low-cost DNA synthesis, advanced genome editing techniques, and laboratory automation, a plethora of MGE methodologies were recently developed and applied in fields ranging from basic research to applied sectors. This review analyzes one-step and iterative MGE methodologies, with an emphasis on recombineering and CRISPR-Cas systems, and showcases emerging paradigm-shifting applications in biomanufacturing, agriculture, and therapeutics. We conclude by analyzing the limitations of existing technologies and discussing future directions for further optimizing MGE to solve system-level problems.
