Applications of piggyBac Transposons for Genome Manipulation in Stem Cells

Abstract

Transposons are mobile genetic elements in the genome. The piggyBac (PB) transposon system is increasingly being used for stem cell research due to its high transposition efficiency and seamless excision capacity. Over the past few decades, forward genetic screens based on PB transposons have been successfully established to identify genes associated with drug resistance and stem cell-related characteristics. Moreover, PB transposon is regarded as a promising gene therapy vector and has been used in some clinically relevant stem cells. Here, we review the recent progress on the basic biology of PB, highlight its applications in current stem cell research, and discuss its advantages and challenges.

Establishment of a novel minigenome system for the identification of drugs targeting Nipah virus replication

“…However, the length of the NiV-L gene was 6735 bp, which exceeds the cargo limit of the lentiviral vectors and will lead to a lower packaging efficiency of the lentivirus. Therefore, we use the piggyBac system, which can transfer large genes (>100 kb) to target cells [14], for NiV-L gene delivery. After successful lentiviral transduction and transposition, BHK-PNL helper cells were established following continuous selection using G418, hygromycin B and blasticidin S (Fig.…”

Protocol for inducible piggyBac transposon system for efficient gene overexpression in human pluripotent stem cells

“…Compared to the other methods to overexpress of specific genes, such as plasmid DNA transfection, retroviral- or lentiviral-based infection, electroporation, the PiggyBac transposon system is technically simpler and higher efficiency, it has a large cargo capacity (up to 10 KB) with multiple genes expression at the same time, and the donor cassette can be excised using excision-only PiggyBac transposase in scarless fashion if we do not need them later. PiggyBac transposon system is broadly used tool that allows DNA cargos inserted into genome in “AATT” sequence with high efficiency, which has been applied in gene overexpression, knockdown, genome editing in various mammalian cells, including stem cell ( Schertzer et al., 2019 ; Sun et al., 2021 ). However, we did not perform the gene overexpression in other stem cells, such as cancer stem cells.…”