piggyBac-Based Non-Viral In Vivo Gene Delivery Useful for Production of Genetically Modified Animals and Organs

Abstract

In vivo gene delivery involves direct injection of nucleic acids (NAs) into tissues, organs, or tail-veins. It has been recognized as a useful tool for evaluating the function of a gene of interest (GOI), creating models for human disease and basic research targeting gene therapy. Cargo frequently used for gene delivery are largely divided into viral and non-viral vectors. Viral vectors have strong infectious activity and do not require the use of instruments or reagents helpful for gene delivery but bear immunological and tumorigenic problems. In contrast, non-viral vectors strictly require instruments (i.e., electroporator) or reagents (i.e., liposomes) for enhanced uptake of NAs by cells and are often accompanied by weak transfection activity, with less immunological and tumorigenic problems. Chromosomal integration of GOI-bearing transgenes would be ideal for achieving long-term expression of GOI. piggyBac (PB), one of three transposons (PB, Sleeping Beauty (SB), and Tol2) found thus far, has been used for efficient transfection of GOI in various mammalian cells in vitro and in vivo. In this review, we outline recent achievements of PB-based production of genetically modified animals and organs and will provide some experimental concepts using this system.

Evaluation of Nonviral piggyBac and lentiviral Vector in Functions of CD19chimeric Antigen Receptor T Cells and Their Antitumor Activity for CD19+ Tumor Cells

“…Viral vectors include adenoviral (AV), adeno-associated viral (AAV), and lentiviral (LV) vectors. AV vectors have immunogenic properties when directly administered in vivo , whereas LV vectors require large-scale production by living cells ( 4 ). Nonviral delivery systems include cationic lipids and cationic polymers ( 5 ), as well as nonviral plasmids.…”

“…Transposons are natural vectors for gene delivery ( 6 ). These vectors, which carry a gene expression unit called a “transgene,” can overcome some of the limitations of viral vectors ( 4 ). Three transposons have been described to date, sleeping beauty (SB), Tol2 , and piggyback (PB).…”

A native, highly activeTc1/marinertransposon from zebrafish (ZB) offers an efficient genetic manipulation tool for vertebrates

“…However, only Tol2 has been widely used as a genetic tool, particularly in the zebrafish model ( 21 , 22 ). The PB transposon is a member of the piggyBac transposon superfamily present in the genomes of a wide range of organisms, including fungi, plants, insects, crustaceans, urochordates, amphibians, fishes and mammals ( 23 , 24 ). In addition, putatively active PB -like transposons have been identified in moths ( 25 ), silkworms ( 26 ), ants ( 27 ), Xenopus ( 28 ) and bats ( 29 ).…”

In vivo Hepatocyte Genome Manipulation via Intravenous Injection of Genome Editing Components

“…However, recently, sustained transgene expression has become possible when both the piggyBac (PB) transposon vector (carrying a transgene) and PB transposase expression vector are intravenously introduced into mice as naked DNA, or DNA complexed with gene delivery accelerating agents [10][11][12]. The PB gene delivery system is one of the transposons that confer the integration of a target gene into host chromosomes with the aid of transposase activity [13,14]. Therefore, sustained expression of a transgene is thought to be caused by the chromosomal integration of the transgene.…”