Inverted terminal repeats of the TcBuster DNA transposon (TcBuster ITRs)

TcBuster is a DNA transposon belonging to the hAT (hobo-Ac-Tam3) superfamily, originally identified in the red flour beetle Tribolium castaneum (Woodard & Wilson, 2015). The inverted terminal repeats (ITRs) are essential DNA sequences located at the ends of the transposon that serve as the specific recognition sites for the TcBuster transposase enzyme (Hacker et al., 2017). In the field of gene therapy, the TcBuster system is utilized as a non-viral vector for the stable integration of therapeutic genes into the host genome. The transposase recognizes the ITRs and performs a cut-and-paste mechanism, excising the DNA segment between the repeats and inserting it into the target cell's DNA (Ammar et al., 2012). This system is increasingly used in the manufacturing of chimeric antigen receptor (CAR) T-cells due to its ability to carry large genetic payloads and its high integration efficiency in human primary cells (Luminary Therapeutics, 2023). Compared to viral vectors, the TcBuster system offers potential advantages in terms of reduced manufacturing costs and improved safety profiles regarding immunogenicity. However, like all integrating vectors, it carries risks of insertional mutagenesis and requires careful monitoring of integration sites (Gogol-Döring et al., 2016). The ITRs are the critical structural components that define the boundaries of the genetic material to be transferred.

Other names

TcB ITRsTcBuster terminal repeatsTcBuster inverted repeatsTcBuster DNA recognition sequences

Transposase-mediated cut-and-paste DNA transposition involving excision from a donor plasmid and integration into the host genome.