Improvement in Tol2 transposon for efficient large-cargo capacity transgene applications in cultured cells and zebrafish(Danio rerio)  

作　　者：Peng-Cheng Wang Hao Deng Rang Xu Jiu-Lin Du Rongkun Tao 

机构地区：[1]Department of Pediatric Cardiology,Xinhua Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China [2]Institute of Neuroscience,State Key Laboratory of Neuroscience,Center for Excellence in Brain Science and Intelligence Technology,Chinese Academy of Sciences,Shanghai 200031,China [3]Medical School,Kunming University of Science and Technology,Kunming,Yunnan 650500,China [4]Scientific Research Center,Xinhua Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China [5]University of Chinese Academy of Sciences,Beijing 100049,China [6]School of Life Science and Technology,ShanghaiTech University,Shanghai 200031,China

出　　处：《Zoological Research》2024年第3期567-574,共8页动物学研究（英文）

基　　金：supported by the National Science and Technology Innovation 2030 Major Projects(2021ZD0202200);National Natural Science Foundation of China(32171090,81970264);Shanghai Science and Technology Commission(21ZR1482600);2023 Youth Innovation Promotion Association CAS。

摘　　要：Most viruses and transposons serve as effective carriers for the introduction of foreign DNA up to 11 kb into vertebrate genomes.However,their activity markedly diminishes with payloads exceeding 11 kb.Expanding the payload capacity of transposons could facilitate more sophisticated cargo designs,improving the regulation of expression and minimizing mutagenic risks associated with molecular therapeutics,metabolic engineering,and transgenic animal production.In this study,we improved the Tol2 transposon by increasing protein expression levels using a translational enhancer(QBI SP163,ST)and enhanced the nuclear targeting ability using the nuclear localization protein H2B(SHT).The modified Tol2 and ST transposon efficiently integrated large DNA cargos into human cell cultures(H1299),comparable to the well-established super PiggyBac system.Furthermore,mRNA from ST and SHT showed a significant increase in transgene delivery efficiency of large DNA payloads(8 kb,14 kb,and 24 kb)into zebrafish(Danio rerio).This study presents a modified Tol2 transposon as an enhanced nonviral vector for the delivery of large DNA payloads in transgenic applications.

关 键 词：ZEBRAFISH Tol2 transposase TRANSGENE Large payload Synthetic biology 

分 类 号：Q78[生物学—分子生物学]