Volume 34, No 4, Apr 2024

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 34 Issue 4, April 2024: 309-322   |  Open Access

ORIGINAL ARTICLES

Convenient synthesis and delivery of a megabase-scale designer accessory chromosome empower biosynthetic capacity

Yuan Ma^1,2 , Shuxin Su^1,2 , Zongheng Fu^1,2 , Chu Zhou^1,2 , Bin Qiao¹ , Yi Wu^1,2,* , Ying-Jin Yuan^1,2,*

¹Frontiers Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin, China
²Frontiers Research Institute for Synthetic Biology, Tianjin University, Tianjin, China
Correspondence: Yi Wu(yi.wu@tju.edu.cn)Ying-Jin Yuan(yjyuan@tju.edu.cn)

Synthetic biology confers new functions to hosts by introducing exogenous genetic elements, yet rebuilding complex traits that are based on large-scale genetic information remains challenging. Here, we developed a CRISPR/Cas9-mediated haploidization method that bypasses the natural process of meiosis. Based on the programmed haploidization in yeast, we further developed an easy-to-use method designated HAnDy (Haploidization-based DNA Assembly and Delivery in yeast) that enables efficient assembly and delivery of large DNA, with no need for any fussy in vitro manipulations. Using HAnDy, a de novo designed 1.024 Mb synthetic accessory chromosome (synAC) encoding 542 exogenous genes was parallelly assembled and then directly transferred to six phylogenetically diverse yeasts. The synAC significantly promotes hosts’ adaptations and increases the scope of the metabolic network, which allows the emergence of valuable compounds. Our approach should facilitate the assembly and delivery of large-scale DNA for expanding and deciphering complex biological functions.

https://doi.org/10.1038/s41422-024-00934-3

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