Tol2 Transposon-based System Services
Many transgenic tools have been adapted from other organisms and optimized for the application of zebrafish to allow stable protein expression and the conditional targeting of proteins for expression in specified tissues. DNA transposons are effective chromosomal engineering vehicles for generating transgenic zebrafish. One of the most popular systems for making transgenic animals in zebrafish is the Tol2 transposon system.
The Tol2 transposon belongs to the hAT family, cloned from Oryzias latipes. In order to adapt Tol2 for use as a gene delivery vehicle, the transposase coding sequence was stripped from the transposon and replaced with a promoter and transgene. mRNA encoding the transposase is provided in trans, guaranteeing that the engineered transposon does not relocate in the genome.
Features of Tol2 Transposon System
- The integration of the targeted gene by Tol2 is mediated by a 'cut-and-paste' mechanism, which does not lead to any rearrangement or modification at the target site except for the creation of an 8-base pair duplication.
- Tol2 transposition efficiency does not significantly decrease with transposon size (up to 10 kb), and it is not sensitive to overexpression inhibition.
- Tol2 transposon shows no preferences with respect to the integration sites which indicates theoretically it can translocate to anywhere on a chromosome.
- Single copy insertions can be achieved and DNA inserts of quite large sizes (up to 13 kilobases) can be cloned between these sequences.
At Creative Biogene, this tol2 transposon-based system has been successfully used to create transgenic zebrafish with large fragment insertion and high efficiency. By our tol2 transgenesis services you can generate:
- Transcription reporters
- Cre Recombinase-expressing lines
- Zebrafish models to control the timing of gene expression
- Zebrafish models to analyze tissue-specific gene expression
- Humanized zebrafish
The Tol2 containing vector is capable of accommodating the DNA inserts of up to 10 kb. Through injecting a foreign DNA cloned into a Tol2-transposon vector together with the transposase mRNA into the cell of the one-cell stage embryos, we can obtain the Founder zebrafish easily. Please feel free to contact us or send us an inquiry for more information or a formal quote.
References
- Juntti S A, et al. Tol2-mediated generation of a transgenic haplochromine cichlid, Astatotilapia burtoni. PLoS One, 2013, 8(10).
- Clark K J, et al. Transgenic zebrafish using transposable elements. Methods in cell biology. Academic Press, 2011, 104: 137-149.
- Ni J, et al. Active recombinant Tol2 transposase for gene transfer and gene discovery applications. Mobile DNA, 2016, 7(1): 6.
- Elena Kardash. Zebrafish Research Methods. MATER METHODS. 2012;2:109.
For research use only. Not intended for any clinical use.
