Excision-only PiggyBac Transposase Expression Vector

Remove a PiggyBac insert from the genome and restore the original genomic sequence with this Excision-only PiggyBac Transposase
  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers

Products

Catalog Number Description Size Price Quantity Add to Cart
PB220PA-1 Excision only piggyBac Transposase expression vector 10 µg $462
- +

Overview

Overview

Reversible transgenesis that leaves no trace behind

One of the advantages of the PiggyBac Transposon System* is the reversibility of the integration event. With the Excision-only PiggyBac Transposase, delivered by transient transfection of the Exicison-only PiggyBac Transposase Expression Vector, you can remove any DNA integrated into the genome using the PiggyBac Transposon System. After removal of the PiggyBac insert, the genomic DNA is restored to the original sequence with no residual PiggyBac sequences left behind, resulting in a truly footprint-free removal.

Both the Excision-only PiggyBac Transposase and the Super PiggyBac Transposase (Cat.# PB210PA-1) recognize transposon-specific inverted terminal repeats (ITRs). While the Super PiggyBac Transposase has both excision and integration activities that enable the enzyme to mediate removal of the ITRs and intervening DNA from one location (the vector) and insert that DNA fragment into another location (the genome), the Excision-only PiggyBac Transposase only has the excision activity. Thus, the Excision-only PiggyBac Transposase can be used to remove DNA segments from the genome that are bordered by the PiggyBac transposon ITRs.

With the PiggyBac Transposon System, you can:

  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers
  • Determine the number of integration events with the PiggyBac qPCR Copy Number Kit (# PBC100A-1)

Customer Agreements

Academic customers can purchase PiggyBac Transposon System components for internal research purposes for indefinite use, whereas commercial customers must sign a customer agreement for a four-month, limited-use license to evaluate the technology.

For end user license information, see the following:

* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, Inc.

How It Works

How It Works

The PiggyBac Transposon System’s Cut-and-Paste Mechanism

The efficient PiggyBac Transposon System uses a cut-and-paste mechanism to transfer DNA from the PiggyBac Vector into the genome. If only temporary genomic integration is desired, the Excision-only PiggyBac Transposase can be transiently expressed for footprint-free removal of the insert, resulting in reconstitution of the original genome sequence.

The PiggyBac Transposon System’s cut-and-paste mechanism

Figure 1. The PiggyBac Transposon System’s cut-and-paste mechanism.

  • The Super PiggyBac Transposase binds to specific inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excises the ITRs and intervening DNA.
  • The Super PiggyBac Transposase inserts the ITR-Expression Cassette-ITR segment into the genome at TTAA sites.
  • The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassette-ITR segment from the genome, for footprint-free removal

Supporting Data

Supporting Data

Reverse transgenesis with the PiggyBac Transposon System’s footprint-free excision process

SBI’s Excision-only PiggyBac Transposase is integrase-deficient but excision capable for reversible transgenesis

Figure 2. SBI’s Excision-only PiggyBac Transposase is integrase-deficient but excision capable for reversible transgenesis. (Left panels) Comparison of the integration abilities of the Super PiggyBac Transposase (Cat.# PB210PA-1) to the Excision-only PiggyBac Transposase upon co-transfection with a GFP-expressing PiggyBac Vector. The very low number of GFP-positive cells in the sample co-transfected with Excision-only Transposase versus the numerous GFP-positive cells in the sample co-transfected with the Super PiggyBac Transposase demonstrate the poor integration abilities of the Excision-only enzyme. (Right panels) We inserted a GFP expression cassette into the genome, and then inactivated GFP expression by disrupting the open reading frame with a PiggyBac transposon. Successful use of the Excision-only PiggyBac Transposase restores GFP function, leading to GFP-positive cells. Restoration of GFP fluorescence also demonstrates the seamlessness of excision—the GFP coding region would remain disrupted if the transposon sequences did not get completely removed.

Resources

Citations

  • Uchino, S, et al. (2022) Live imaging of transcription sites using an elongating RNA polymerase II-specific probe. The Journal of cell biology. 1970 Jan 1; 221(2). PM ID: 34854870
  • Teixeira, A, et al. (2022) CelloSelect – A synthetic cellobiose metabolic pathway for selection of stable transgenic CHO cell lines. Metabolic Engineering. 1970 Jan 1; 70:23-30. Link: Metabolic Engineering
  • Rui, Y, et al. (2022) High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Science advances. 1970 Jan 1; 8(1):eabk2855. PM ID: 34985952
  • Vásquez-Limeta, A, et al. (2022) CPAP insufficiency leads to incomplete centrioles that duplicate but fragment. The Journal of cell biology. 1970 Jan 1; 221(5). PM ID: 35404385
  • Su, CJ, et al. (2022) Ligand-receptor promiscuity enables cellular addressing. Cell systems. 1970 Jan 1;. PM ID: 35421362
  • Klumpe, HE, et al. (2022) The context-dependent, combinatorial logic of BMP signaling. Cell systems. 1970 Jan 1;. PM ID: 35421361
  • Kitano, H, Kawabe, Y & Kamihira, M. (2022) HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver Functions. Cells. 1970 Jan 1; 11(7). PM ID: 35406758
  • Sugiman-Marangos, SN, et al. (2022) Structures of distant diphtheria toxin homologs reveal functional determinants of an evolutionarily conserved toxin scaffold. Communications biology. 1970 Jan 1; 5(1):375. PM ID: 35440624
  • Ma, X, et al. (2022) Validation of reliable safe harbor locus for efficient porcine transgenesis. Functional & integrative genomics. 1970 Jan 1;. PM ID: 35412198
  • Nishimura, K, et al. (2022) Rapid conversion of human induced pluripotent stem cells into dopaminergic neurons by inducible expression of two transcription factors. Stem cells and development. 1970 Jan 1;. PM ID: 35420042
  • Liu, Z, Ramirez, A & Liu, X. (2022) Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2488:227-236. PM ID: 35347692
  • Dandridge, S. (2022) Honors Thesis: Defining the effect of zinc on the proliferation of MDA-MB-231 cells compared to MCF10A cells. Thesis. 1970 Jan 1;. Link: Thesis
  • Yang, D, et al. (2022) Lineage tracing reveals the phylodynamics, plasticity, and paths of tumor evolution. Cell. 1970 Jan 1; 185(11):1905-1923.e25. PM ID: 35523183
  • Biswas, S, et al. (2022) Long-term hepatitis B virus infection of rhesus macaques requires suppression of host immunity. Nature communications. 1970 Jan 1; 13(1):2995. PM ID: 35637225
  • Breau, KA, et al. (2022) Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells. Stem cell reports. 1970 Jan 1;. PM ID: 35523179
  • Lensch, S, et al. (2022) Dynamic spreading of chromatin-mediated gene silencing and reactivation between neighboring genes in single cells. eLife. 1970 Jan 1; 11. PM ID: 35678392
  • Gu, J, Sumer, H & Cromer, B. (2022) Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2495:49-66. PM ID: 35696027
  • Wang, S, et al. (2021) Budding epithelial morphogenesis driven by cell-matrix versus cell-cell adhesion. Cell. 1970 Jan 1;. PM ID: 34133940
  • Ng, YH, et al. (2021) Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem cell reports. 1970 Jan 1;. PM ID: 34171286
  • Ukaji, T, et al. (2021) Novel knock-in mouse model for the evaluation of the therapeutic efficacy and toxicity of human podoplanin-targeting agents. Cancer science. 1970 Jan 1; 112(6):2299-2313. PM ID: 33735501

Products

Catalog Number Description Size Price Quantity Add to Cart
PB220PA-1 Excision only piggyBac Transposase expression vector 10 µg $462
- +

Overview

Overview

Reversible transgenesis that leaves no trace behind

One of the advantages of the PiggyBac Transposon System* is the reversibility of the integration event. With the Excision-only PiggyBac Transposase, delivered by transient transfection of the Exicison-only PiggyBac Transposase Expression Vector, you can remove any DNA integrated into the genome using the PiggyBac Transposon System. After removal of the PiggyBac insert, the genomic DNA is restored to the original sequence with no residual PiggyBac sequences left behind, resulting in a truly footprint-free removal.

Both the Excision-only PiggyBac Transposase and the Super PiggyBac Transposase (Cat.# PB210PA-1) recognize transposon-specific inverted terminal repeats (ITRs). While the Super PiggyBac Transposase has both excision and integration activities that enable the enzyme to mediate removal of the ITRs and intervening DNA from one location (the vector) and insert that DNA fragment into another location (the genome), the Excision-only PiggyBac Transposase only has the excision activity. Thus, the Excision-only PiggyBac Transposase can be used to remove DNA segments from the genome that are bordered by the PiggyBac transposon ITRs.

With the PiggyBac Transposon System, you can:

  • Make transgenic cell lines with a single transfection
  • Integrate multiple PiggyBac Vectors in a single transfection
  • Insert an expression cassette into human, mouse, and rat cells
  • Deliver virtually any-sized DNA insert, from 10 – 100 kb
  • Choose from PiggyBac Vectors that express your gene-of-interest from constitutive or inducible promoters and include a variety of markers
  • Determine the number of integration events with the PiggyBac qPCR Copy Number Kit (# PBC100A-1)

Customer Agreements

Academic customers can purchase PiggyBac Transposon System components for internal research purposes for indefinite use, whereas commercial customers must sign a customer agreement for a four-month, limited-use license to evaluate the technology.

For end user license information, see the following:

* SBI is fully licensed to distribute PiggyBac vectors as a partnership with Hera BioLabs, Inc.

How It Works

How It Works

The PiggyBac Transposon System’s Cut-and-Paste Mechanism

The efficient PiggyBac Transposon System uses a cut-and-paste mechanism to transfer DNA from the PiggyBac Vector into the genome. If only temporary genomic integration is desired, the Excision-only PiggyBac Transposase can be transiently expressed for footprint-free removal of the insert, resulting in reconstitution of the original genome sequence.

The PiggyBac Transposon System’s cut-and-paste mechanism

Figure 1. The PiggyBac Transposon System’s cut-and-paste mechanism.

  • The Super PiggyBac Transposase binds to specific inverted terminal repeats (ITRs) in the PiggyBac Cloning and Expression Vector and excises the ITRs and intervening DNA.
  • The Super PiggyBac Transposase inserts the ITR-Expression Cassette-ITR segment into the genome at TTAA sites.
  • The Excision-only Super PiggyBac Transposase can be used to remove the ITR-Expression Cassette-ITR segment from the genome, for footprint-free removal

Supporting Data

Supporting Data

Reverse transgenesis with the PiggyBac Transposon System’s footprint-free excision process

SBI’s Excision-only PiggyBac Transposase is integrase-deficient but excision capable for reversible transgenesis

Figure 2. SBI’s Excision-only PiggyBac Transposase is integrase-deficient but excision capable for reversible transgenesis. (Left panels) Comparison of the integration abilities of the Super PiggyBac Transposase (Cat.# PB210PA-1) to the Excision-only PiggyBac Transposase upon co-transfection with a GFP-expressing PiggyBac Vector. The very low number of GFP-positive cells in the sample co-transfected with Excision-only Transposase versus the numerous GFP-positive cells in the sample co-transfected with the Super PiggyBac Transposase demonstrate the poor integration abilities of the Excision-only enzyme. (Right panels) We inserted a GFP expression cassette into the genome, and then inactivated GFP expression by disrupting the open reading frame with a PiggyBac transposon. Successful use of the Excision-only PiggyBac Transposase restores GFP function, leading to GFP-positive cells. Restoration of GFP fluorescence also demonstrates the seamlessness of excision—the GFP coding region would remain disrupted if the transposon sequences did not get completely removed.

Citations

  • Uchino, S, et al. (2022) Live imaging of transcription sites using an elongating RNA polymerase II-specific probe. The Journal of cell biology. 1970 Jan 1; 221(2). PM ID: 34854870
  • Teixeira, A, et al. (2022) CelloSelect – A synthetic cellobiose metabolic pathway for selection of stable transgenic CHO cell lines. Metabolic Engineering. 1970 Jan 1; 70:23-30. Link: Metabolic Engineering
  • Rui, Y, et al. (2022) High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Science advances. 1970 Jan 1; 8(1):eabk2855. PM ID: 34985952
  • Vásquez-Limeta, A, et al. (2022) CPAP insufficiency leads to incomplete centrioles that duplicate but fragment. The Journal of cell biology. 1970 Jan 1; 221(5). PM ID: 35404385
  • Su, CJ, et al. (2022) Ligand-receptor promiscuity enables cellular addressing. Cell systems. 1970 Jan 1;. PM ID: 35421362
  • Klumpe, HE, et al. (2022) The context-dependent, combinatorial logic of BMP signaling. Cell systems. 1970 Jan 1;. PM ID: 35421361
  • Kitano, H, Kawabe, Y & Kamihira, M. (2022) HepG2-Based Designer Cells with Heat-Inducible Enhanced Liver Functions. Cells. 1970 Jan 1; 11(7). PM ID: 35406758
  • Sugiman-Marangos, SN, et al. (2022) Structures of distant diphtheria toxin homologs reveal functional determinants of an evolutionarily conserved toxin scaffold. Communications biology. 1970 Jan 1; 5(1):375. PM ID: 35440624
  • Ma, X, et al. (2022) Validation of reliable safe harbor locus for efficient porcine transgenesis. Functional & integrative genomics. 1970 Jan 1;. PM ID: 35412198
  • Nishimura, K, et al. (2022) Rapid conversion of human induced pluripotent stem cells into dopaminergic neurons by inducible expression of two transcription factors. Stem cells and development. 1970 Jan 1;. PM ID: 35420042
  • Liu, Z, Ramirez, A & Liu, X. (2022) Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2488:227-236. PM ID: 35347692
  • Dandridge, S. (2022) Honors Thesis: Defining the effect of zinc on the proliferation of MDA-MB-231 cells compared to MCF10A cells. Thesis. 1970 Jan 1;. Link: Thesis
  • Yang, D, et al. (2022) Lineage tracing reveals the phylodynamics, plasticity, and paths of tumor evolution. Cell. 1970 Jan 1; 185(11):1905-1923.e25. PM ID: 35523183
  • Biswas, S, et al. (2022) Long-term hepatitis B virus infection of rhesus macaques requires suppression of host immunity. Nature communications. 1970 Jan 1; 13(1):2995. PM ID: 35637225
  • Breau, KA, et al. (2022) Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells. Stem cell reports. 1970 Jan 1;. PM ID: 35523179
  • Lensch, S, et al. (2022) Dynamic spreading of chromatin-mediated gene silencing and reactivation between neighboring genes in single cells. eLife. 1970 Jan 1; 11. PM ID: 35678392
  • Gu, J, Sumer, H & Cromer, B. (2022) Efficient Generation of Stable Cell Lines with Inducible Neuronal Transgene Expression Using the piggyBac Transposon System. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2495:49-66. PM ID: 35696027
  • Wang, S, et al. (2021) Budding epithelial morphogenesis driven by cell-matrix versus cell-cell adhesion. Cell. 1970 Jan 1;. PM ID: 34133940
  • Ng, YH, et al. (2021) Efficient generation of dopaminergic induced neuronal cells with midbrain characteristics. Stem cell reports. 1970 Jan 1;. PM ID: 34171286
  • Ukaji, T, et al. (2021) Novel knock-in mouse model for the evaluation of the therapeutic efficacy and toxicity of human podoplanin-targeting agents. Cancer science. 1970 Jan 1; 112(6):2299-2313. PM ID: 33735501