PUBLICATION
Improvement in Tol2 transposon for efficient large-cargo capacity transgene applications in cultured cells and zebrafish ( Danio rerio)
- Authors
- Wang, P.C., Deng, H., Xu, R., Du, J.L., Tao, R.
- ID
- ZDB-PUB-240517-12
- Date
- 2024
- Source
- Zoological research 45: 567574567-574 (Journal)
- Registered Authors
- Du, Jiu Lin
- Keywords
- Large payload, Synthetic biology, Tol2 transposase, Transgene, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- DNA Transposable Elements*/genetics
- Gene Transfer Techniques
- Humans
- Transgenes*
- Zebrafish*/genetics
- PubMed
- 38757224 Full text @ Zool Res
Citation
Wang, P.C., Deng, H., Xu, R., Du, J.L., Tao, R. (2024) Improvement in Tol2 transposon for efficient large-cargo capacity transgene applications in cultured cells and zebrafish ( Danio rerio). Zoological research. 45:567574567-574.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping