Abstract
Sleeping Beauty (SB) is a transposon system that has been widely used as a genetic engineering tool. Central to the development of any transposon as a research tool is the ability to integrate a foreign piece of DNA into the cellular genome. Driven by the need for efficient transposon-based gene vector systems, extensive studies have largely elucidated the molecular actors and actions taking place during SB transposition. Close transposon relatives and other recombination enzymes, including retroviral integrases, have served as useful models to infer functional information relevant to SB. Recently obtained structural data on the SB transposase enable a direct insight into the workings of this enzyme. These efforts cumulatively allowed the development of novel variants of SB that offer advanced possibilities for genetic engineering due to their hyperactivity, integration deficiency, or targeting capacity. However, many aspects of the process of transposition remain poorly understood and require further investigation. We anticipate that continued investigations into the structure–function relationships of SB transposition will enable the development of new generations of transposition-based vector systems, thereby facilitating the use of SB in preclinical studies and clinical trials.
Highlights
The capacity of nucleic acids to move around and integrate into a new locus has evolved in manifold ways
Many autonomous Transposable elements (TEs) have given rise to non-autonomous derivatives by mutations, insertions, or deletions in their transposase coding regions. These non-autonomous TEs can still be mobilized, but need a functional transposase expressed by another element in the same cell [14]. It is this trans-complementarity between two functional components that serves as the basis of turning transposons into genetic vector systems suitable for moving any gene of interest into the genome of a host cell
We described the structural features and mechanistic steps involved in Sleeping Beauty (SB) transposition
Summary
The capacity of nucleic acids to move around and integrate into a new locus has evolved in manifold ways. Class I TEs, called retrotransposons, follow a copy-and-paste mechanism After transcription of their DNA genome to RNA, a reverse transcription step back into DNA is performed, and a reintegration into the genome occurs [6]. Subclass II transposons, such as members of the Helitron superfamily [9], follow a copy-and-paste mechanism, during which the element generates copies of itself which integrate into the genome. These non-autonomous TEs can still be mobilized, but need a functional transposase expressed by another element in the same cell [14] It is this trans-complementarity between two functional components (the transposase and the specific TIRs that are recognized and mobilized by the transposase) that serves as the basis of turning transposons into genetic vector systems suitable for moving any gene of interest into the genome of a host cell. The structural features and mechanistic steps and processes taking place in the life cycle of SB from DNA binding up to integration are described
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