The piggyBac transposase technology is a non-viral, gene delivery system designed for use in many different research applications. With a large cargo capacity of over 200kb, piggyBac surpasses many other transposon and viral delivery vehicles, making it the optimal choice for stable cell line creation. The high efficiency and stability of piggyBac integration events have proven effective in a variety of genomes. To date, over 400 peer-reviewed papers have been published as a result of the piggyBac technology. Hera provides piggyBac products & licenses, which include research and commercial applications for drug discovery & early development.
How piggyBac works
How it works
Step 1: Clone cargo into transposon vector. Step 2: Co-transfect Transposon and Transposase into cells.
Step 3: Transposase cuts out cargo and pastes into random TTAA sites.
Step 4: Screen clones for desired insert.
Step 5 (optional): Use Excision-only piggyBac for Footprint-Free gene editing
Features
Small to enormous gene integration (200KB+)
Very efficient
High expression
Stable, seamless removal if desired
piggyBac outperforms Leap-In transposase
Approximately 4 to 11-fold greater titers compared to conventional gene integration systems for monoclonal antibodies (mAbs) or bi- specific antibodies (BSAb)1
Only 1.4 to 1.7 greater titers compared to conventional with Leap-In2
1: Rajendra et al. (2016) Generation of Stable Chinese Hamster Ovary Pools Yielding Antibody Titers of up to 7.6 g/L Using the piggyBac Transposon System. Biotechnol. Prog., Vol. 32
2: Balasubramanian et al. (2018) Generation of high expressing Chinese Hamster Ovary cell pools using the Leap-In transposon system. Biotechnol J. 2018 Oct;13(10):e1700748
PiggyBac generated luciferase reporter cells lines show enhanced stability.
Mossine VV, Waters JK, Hannink M, Mawhinney TP (2013) piggyBac Transposon plus Insulators Overcome Epigenetic Silencing to Provide for Stable Signaling Pathway Reporter Cell Lines. PLoS ONE 8(12): e85494. doi:10.1371/journal.pone.0085494
Creation of BAC transgenic SRIPA rats with piggyBac compared to CRISPR/TALEN nucleases.
Jung, C. J., Ménoret, S., Brusselle, L., Tesson, L., Usal, C., Chenouard, V., . . . Anegon, I. (2016). Comparative Analysis of piggyBac, CRISPR/Cas9 and TALEN Mediated BAC Transgenesis in the Zygote for the Generation of Humanized SIRPA Rats. Sci. Rep. Scientific Reports, 6, 31455. doi:10.1038/srep31455
PiggyBac Reporter Hep-G2 cell line for high throughput and high sensitivity screening of AhR transcription and study of cytotoxic compounds.
Novotna, A., Petr, P., & Dvorak, Z. (2011). Novel Stably Transfected Gene Reporter Human Hepatoma Cell Line for Assessment of Aryl Hydrocarbon Receptor Transcriptional Activity: Construction and Characterization. Environmental Science & Technology Environ. Sci. Technol., 45(23), 10133-10139. doi:10.1021/es2029334- Generation of Stable Chinese Hamster Ovary Pools Yielding Antibody Titers of up to 7.6 g/L Using the piggyBac Transposon System
Rajendra et al. Eli Lilly (2016) Biotechnol. Prog. Vol. 32, No. 5