Efficient and multiplex gene upregulation in plants through CRISPR-Cas-mediated knockin of enhancers

Yao, Qi1,2,3; Shen, Rundong2; Shao, Yang4; Tian, Yifu2; Han, Peijin2; Zhang, Xuening2; Zhu, Jian-Kang5,6; Lu, Yuming1

2024-09-02

10.1016/j.molp.2024.07.009

MOLECULAR PLANT   影响因子和分区

1674-2052

1752-9867

Gene upregulation through genome editing is important for plant research and breeding. Targeted insertion of short transcriptional enhancers (STEs) into gene promoters may offer a universal solution akin to transgene-mediated overexpression while avoiding the drawbacks associated with transgenesis. Here, we introduce an "in locus activation"technique in rice that leverages well-characterized STEs for refined, heritable, and multiplexed gene upregulation. To address the scarcity of potent enhancers, we developed a large-scale mining approach and discovered a suite of STEs that are capable of enhancing gene expression in rice protoplasts. The in locus integration of these STEs into eight rice genes resulted in substantial transcriptional upregulation in the edited plants, with up to 869.1-fold increases in their transcript levels. Employing a variety of STEs, we achieved delicate control of gene expression, enabling the fine-tuning of key phenotypic traits such as plant height. Our approach also enabled efficient multiplexed gene upregulation, with up to four genes activated simultaneously, significantly enhancing the nicotinamide mononucleotide metabolic pathway. Importantly, heritability studies from the T0 to T3 generations confirmed the stable and heritable nature of STE-driven gene activation. Collectively, our work demonstrates that coupled with STE mining, leveraging genome editing for in locus activation and gene upregulation holds great promise to be widely adopted in fundamental plant research and crop breeding.

genome editing short transcriptional enhancers gene upregulation crop breeding

SCI

英语

通讯

National Key R&D Program of China["2021YFD1201300","2021YFA1300404"] ; Shanghai Agricultural Science and Technology Innovation Program of China[K2023001] ; National Natural Science Foundation of China["32070396","32188102"]

Biochemistry & Molecular Biology ; Plant Sciences

Biochemistry & Molecular Biology ; Plant Sciences

WOS:001317215500001

CELL PRESS

Web of Science

1.Shanghai Jiao Tong Univ, Shanghai Collaborat Innovat Ctr Agriseeds, Joint Ctr Single Cell Biol, Sch Agr & Biol, Shanghai 200240, Peoples R China
2.Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, Ctr Excellence Mol Plant Sci, Shanghai 201602, Peoples R China
3.Univ Chinese Acad Sci, Beijing, Peoples R China
4.Huazhong Agr Univ, Coll Plant Sci & Technol, Wuhan 430070, Peoples R China
5.Southern Univ Sci & Technol, Inst Adv Biotechnol, Shenzhen 518055, Peoples R China
6.Southern Univ Sci & Technol, Sch Med, Shenzhen 518055, Peoples R China

Yao, Qi,Shen, Rundong,Shao, Yang,et al. Efficient and multiplex gene upregulation in plants through CRISPR-Cas-mediated knockin of enhancers[J]. MOLECULAR PLANT,2024,17(9).

Yao, Qi.,Shen, Rundong.,Shao, Yang.,Tian, Yifu.,Han, Peijin.,...&Lu, Yuming.(2024).Efficient and multiplex gene upregulation in plants through CRISPR-Cas-mediated knockin of enhancers.MOLECULAR PLANT,17(9).

Yao, Qi,et al."Efficient and multiplex gene upregulation in plants through CRISPR-Cas-mediated knockin of enhancers".MOLECULAR PLANT 17.9(2024).