دورية أكاديمية
Improvement in Tol2 transposon for efficient large-cargo capacity transgene applications in cultured cells and zebrafish ( Danio rerio ).
| العنوان: | Improvement in Tol2 transposon for efficient large-cargo capacity transgene applications in cultured cells and zebrafish ( Danio rerio ). |
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| المؤلفون: | Wang PC; Department of Pediatric Cardiology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China.; Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China., Deng H; Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.; Medical School, Kunming University of Science and Technology, Kunming, Yunnan 650500, China., Xu R; Scientific Research Center, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China. E-mail: rangxu@shsmu.edu.cn., Du JL; Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.; University of Chinese Academy of Sciences, Beijing 100049, China.; School of Life Science and Technology, ShanghaiTech University, Shanghai 200031, China. E-mail: forestdu@ion.ac.cn., Tao R; Institute of Neuroscience, State Key Laboratory of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China. E-mail: rktao@ion.ac.cn. |
| المصدر: | Zoological research [Zool Res] 2024 May 18; Vol. 45 (3), pp. 567-574. |
| نوع المنشور: | Journal Article; Letter |
| اللغة: | English |
| بيانات الدورية: | Publisher: Science Press Country of Publication: China NLM ID: 101697192 Publication Model: Print Cited Medium: Internet ISSN: 2095-8137 (Print) Linking ISSN: 20958137 NLM ISO Abbreviation: Zool Res Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Beijing, China : Science Press, 2016- |
| مواضيع طبية MeSH: | Zebrafish*/genetics , DNA Transposable Elements*/genetics , Transgenes*, Animals ; Humans ; Animals, Genetically Modified ; Gene Transfer Techniques |
| مستخلص: | 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. |
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| فهرسة مساهمة: | Keywords: Large payload; Synthetic biology; Tol2 transposase; Transgene; Zebrafish Local Abstract: [Publisher, Chinese] 大多数病毒和转座子可将长达11 kb的外源DNA有效引入脊椎动物基因组，但递送超过11 kb的DNA片段时，其活性明显减弱。扩大转座子的有效载荷能力将能够容纳更大的DNA片段、增强调节基因表达的能力，并最大限度地减少分子治疗、代谢工程和转基因动物相关的突变风险。在该研究中，我们通过翻译增强子QBI SP163和核定位蛋白H2B提高蛋白表达水平和细胞核靶向能力，产生了改善的Tol2转座子，ST和SHT。Tol2和ST转座子能有效地将大片段DNA整合到H1299细胞中，其效果可与改善的super PiggyBac转座子系统相媲美。我们发现ST和SHT mRNA能显著提高斑马鱼中大DNA片段（~8 kb， ~14 kb， ~24 kb）的转基因效率。该研究介绍了优化的Tol2转座子可作为一种改进的非病毒载体，应用于递送大DNA片段的转基因实验。. |
| تواريخ الأحداث: | Date Created: 20240517 Date Completed: 20240517 Latest Revision: 20240718 |
| رمز التحديث: | 20240718 |
| مُعرف محوري في PubMed: | PMC11188598 |
| DOI: | 10.24272/j.issn.2095-8137.2024.026 |
| PMID: | 38757224 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2095-8137 |
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| DOI: | 10.24272/j.issn.2095-8137.2024.026 |