PB-EF1α-MCS-IRES-Neo PiggyBac cDNA Cloning and Expression Vector

Easily deliver your gene-of-interest using PiggyBac – this vector uses the EF1α promoter to co-expresses your cDNA and a neomycin marker via an IRES element
  • 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
PB533A-2 PB-EF1α-MCS-IRES-Neo cDNA cloning and expression vector 10 µg $612.00
- +
Contact Us

Overview

Overview

Get easy, consistent transgenesis Consistent and easy-to-use, SBI’s PiggyBac Transposon System includes cloning and expression vectors that come with a range of markers as well as both constitutive and inducible promoters. The PB-EF1α-MCS-IRES-Neo PiggyBac cDNA Cloning and Expression Vector (Cat.# PB533A-2) drives co-expression of your gene-of-interest and the neomycin marker from the moderate EF1α promoter. Co-expression is mediated by an IRES element upstream of the neomycin resistance gene. PB-EF1α-MCS-IRES-Neo PiggyBac cDNA Cloning and Expression VectorWhy use the PiggyBac Transposon System? Easy, consistent transgenesis with no limits on cargo size—For transgenesis that’s easy, consistent, and not limited by cargo size, SBI’s PiggyBac Transposon System is an excellent choice. The system consists of a PiggyBac Vector and the Super PiggyBac Transposase which recognizes transposon-specific inverted terminal repeats (ITRs) and efficiently integrates the ITRs and intervening DNA into the genome at TTAA sites. The Super PiggyBac Transposase is delivered to the cell via the Super PiggyBac Transposase Expression Vector, which is co-transfected with one or more PiggyBac Vectors. Footprint-free removal that leaves no PiggyBac sequences behind—In addition to ease-of-use, consistency, and the lack of limits on DNA insert size, what sets this system apart is the ability to reverse the integration reaction in a footprint-free way—with the Excision Only PiggyBac Transposase (Cat.# PB220PA-1), the ITRs and cargo that the Super PiggyBac Transposase integrates into the genome can be removed, leaving behind the original genomic sequence and nothing else.
  • 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

One transfection can integrate one or more genes that can be precisely removed

Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors

Figure 2. Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors. (Top four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Dual Promoter PiggyBac Cloning and Expression Vector (Cat.# PB513B-1) into HeLa cells demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After ten days of puromycin selection, only the cells co-transfected with the Super PiggyBac Transposase (+PB, right two panels) show robust growth and GFP fluorescence. (Bottom four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Single Promoter PiggyBac Cloning and Expression Vector (Cat.# PB531A-2) into HEK293 cells further demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After seven days of growth, the majority of cells that received the Super PiggyBac Transposase Expression Vector (+PB, right two panels) were RFP positive.

Simultaneous integration of multiple PiggyBac Vectors is also highly efficient

Figure 3. Simultaneous integration of multiple PiggyBac Vectors is also highly efficient. METHODS: Three different PiggyBac transposon vectors (Cat.# PB513B-1, Cat.# PB533A-2, and Cat.# PB531A-2) were co-transfected with (left panels) or without (right panels) the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) into Human HT1080 cells. Puromycin and neomycin selection was applied for seven days. The cells that were co-transfected with the Super PiggyBac Transposase Expression Vector were puro and neo resistant, GFP-positive, and RFP-positive. Background GFP-positive cells that are puro resistant stem from random PB513B-1 integrations during the puromycin selection. The non-PiggyBac-mediated integration rate in those cells was extremely low and no RFP-positive cells were identified.

Resources

Citations

  • 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
  • Ichikawa, M, et al. (2021) Generation of tetracycline-controllable CYP3A4-expressing Caco-2 cells by the piggyBac transposon system. Scientific reports. 1970 Jan 1; 11(1):11670. PM ID: 34083621
  • Sato, Y & Kimura, H. (2021) Dynamic Behavior of Inactive X Chromosome Territory During the Cell Cycle as Revealed by H3K27me3-Specific Intracellular Antibody. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2329:237-247. PM ID: 34085227
  • Kim, JS, Pineda, M & Li, P. (2021) Reconstitution of Morphogen Signaling Gradients in Cultured Cells. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2258:43-56. PM ID: 33340353
  • Thomas, HF, et al. (2021) Temporal dissection of an enhancer cluster reveals distinct temporal and functional contributions of individual elements. Molecular cell. 1970 Jan 1;. PM ID: 33482114
  • Van, MV, Fujimori, T & Bintu, L. (2021) Nanobody-mediated control of gene expression and epigenetic memory. Nature communications. 1970 Jan 1; 12(1):537. PM ID: 33483487
  • Geis, M, et al. (2021) Combinatorial targeting of multiple myeloma by complementing T cell engaging antibody fragments. Communications biology. 1970 Jan 1; 4(1):44. PM ID: 33420283
  • Saijoh, S, et al. (2021) Discovery of a chemical compound that suppresses expression of BEX2, a dormant cancer stem cell-related protein. Biochemical and biophysical research communications. 1970 Jan 1; 537:132-139. PM ID: 33412384
  • Lam, N, Yamanaka, S & Perli, S. (2021) CRISPRi mediated Down regulation of SFPQ Gene Expression in Human induced Pluripotent Stem Cells Results in Massive Cell Death. Research Square. 1970 Jan 1;. Link: Research Square
  • Xiang, K & Bartel, D. (2021) The molecular basis of coupling between poly(A)-tail length and translational efficiency. bioRxiv. 1970 Jan 1;. Link: bioRxiv
  • Sugimoto, S, et al. (2021) An organoid-based organ-repurposing approach to treat short bowel syndrome. Nature. 1970 Jan 1;. PM ID: 33627870
  • Ma, P, et al. (2021) Avidity‐Based Selection of Tissue‐Specific CAR‐T Cells from a Combinatorial Cellular Library of CARs. Advanced Science. 1970 Jan 1;:2003091. Link: Advanced Science
  • Cho, JH, et al. (2021) Engineering advanced logic and distributed computing in human CAR immune cells. Nature communications. 1970 Jan 1; 12(1):792. PM ID: 33542232
  • Omori, S, et al. (2021) Tim4 recognizes carbon nanotubes and mediates phagocytosis leading to granuloma formation. Cell reports. 1970 Jan 1; 34(6):108734. PM ID: 33567275
  • Tjalsma, SJD, et al. (2021) H4K20me1 and H3K27me3 are concurrently loaded onto the inactive X chromosome but dispensable for inducing gene silencing. EMBO reports. 1970 Jan 1;:e51989. PM ID: 33605056
  • Kitano, H, et al. (2021) Development of a genetically modified hepatoma cell line with heat-inducible high liver function. Cytotechnology. 1970 Jan 1;. Link: Cytotechnology
  • Zhu, R, et al. (2021) Synthetic multistability in mammalian cells. bioRxiv. 1970 Jan 1;. Link: bioRxiv
  • Karlsson, J, et al. (2021) Photocrosslinked Bioreducible Polymeric Nanoparticles for Enhanced Systemic siRNA Delivery as Cancer Therapy. Advanced Functional Materials. 1970 Jan 1;:2009768. Link: Advanced Functional Materials
PB-EF1α-MCS-IRES-Neo PiggyBac cDNA Cloning and Expression Vector $612.00

Products

Catalog Number Description Size Price Quantity Add to Cart
PB533A-2 PB-EF1α-MCS-IRES-Neo cDNA cloning and expression vector 10 µg $612.00
- +
Contact Us

Overview

Overview

Get easy, consistent transgenesis Consistent and easy-to-use, SBI’s PiggyBac Transposon System includes cloning and expression vectors that come with a range of markers as well as both constitutive and inducible promoters. The PB-EF1α-MCS-IRES-Neo PiggyBac cDNA Cloning and Expression Vector (Cat.# PB533A-2) drives co-expression of your gene-of-interest and the neomycin marker from the moderate EF1α promoter. Co-expression is mediated by an IRES element upstream of the neomycin resistance gene. PB-EF1α-MCS-IRES-Neo PiggyBac cDNA Cloning and Expression VectorWhy use the PiggyBac Transposon System? Easy, consistent transgenesis with no limits on cargo size—For transgenesis that’s easy, consistent, and not limited by cargo size, SBI’s PiggyBac Transposon System is an excellent choice. The system consists of a PiggyBac Vector and the Super PiggyBac Transposase which recognizes transposon-specific inverted terminal repeats (ITRs) and efficiently integrates the ITRs and intervening DNA into the genome at TTAA sites. The Super PiggyBac Transposase is delivered to the cell via the Super PiggyBac Transposase Expression Vector, which is co-transfected with one or more PiggyBac Vectors. Footprint-free removal that leaves no PiggyBac sequences behind—In addition to ease-of-use, consistency, and the lack of limits on DNA insert size, what sets this system apart is the ability to reverse the integration reaction in a footprint-free way—with the Excision Only PiggyBac Transposase (Cat.# PB220PA-1), the ITRs and cargo that the Super PiggyBac Transposase integrates into the genome can be removed, leaving behind the original genomic sequence and nothing else.
  • 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

One transfection can integrate one or more genes that can be precisely removed

Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors

Figure 2. Efficient transgenesis with the Super PiggyBac Transposase and both single- and dual-promoter PiggyBac Vectors. (Top four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Dual Promoter PiggyBac Cloning and Expression Vector (Cat.# PB513B-1) into HeLa cells demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After ten days of puromycin selection, only the cells co-transfected with the Super PiggyBac Transposase (+PB, right two panels) show robust growth and GFP fluorescence. (Bottom four panels) Co-transfection with the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) and a Single Promoter PiggyBac Cloning and Expression Vector (Cat.# PB531A-2) into HEK293 cells further demonstrates the efficient integration delivered by SBI’s PiggyBac Transposon System. After seven days of growth, the majority of cells that received the Super PiggyBac Transposase Expression Vector (+PB, right two panels) were RFP positive.

Simultaneous integration of multiple PiggyBac Vectors is also highly efficient

Figure 3. Simultaneous integration of multiple PiggyBac Vectors is also highly efficient. METHODS: Three different PiggyBac transposon vectors (Cat.# PB513B-1, Cat.# PB533A-2, and Cat.# PB531A-2) were co-transfected with (left panels) or without (right panels) the Super PiggyBac Transposase Expression Vector (Cat.# PB210PA-1) into Human HT1080 cells. Puromycin and neomycin selection was applied for seven days. The cells that were co-transfected with the Super PiggyBac Transposase Expression Vector were puro and neo resistant, GFP-positive, and RFP-positive. Background GFP-positive cells that are puro resistant stem from random PB513B-1 integrations during the puromycin selection. The non-PiggyBac-mediated integration rate in those cells was extremely low and no RFP-positive cells were identified.

Citations

  • 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
  • Ichikawa, M, et al. (2021) Generation of tetracycline-controllable CYP3A4-expressing Caco-2 cells by the piggyBac transposon system. Scientific reports. 1970 Jan 1; 11(1):11670. PM ID: 34083621
  • Sato, Y & Kimura, H. (2021) Dynamic Behavior of Inactive X Chromosome Territory During the Cell Cycle as Revealed by H3K27me3-Specific Intracellular Antibody. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2329:237-247. PM ID: 34085227
  • Kim, JS, Pineda, M & Li, P. (2021) Reconstitution of Morphogen Signaling Gradients in Cultured Cells. Methods in molecular biology (Clifton, N.J.). 1970 Jan 1; 2258:43-56. PM ID: 33340353
  • Thomas, HF, et al. (2021) Temporal dissection of an enhancer cluster reveals distinct temporal and functional contributions of individual elements. Molecular cell. 1970 Jan 1;. PM ID: 33482114
  • Van, MV, Fujimori, T & Bintu, L. (2021) Nanobody-mediated control of gene expression and epigenetic memory. Nature communications. 1970 Jan 1; 12(1):537. PM ID: 33483487
  • Geis, M, et al. (2021) Combinatorial targeting of multiple myeloma by complementing T cell engaging antibody fragments. Communications biology. 1970 Jan 1; 4(1):44. PM ID: 33420283
  • Saijoh, S, et al. (2021) Discovery of a chemical compound that suppresses expression of BEX2, a dormant cancer stem cell-related protein. Biochemical and biophysical research communications. 1970 Jan 1; 537:132-139. PM ID: 33412384
  • Lam, N, Yamanaka, S & Perli, S. (2021) CRISPRi mediated Down regulation of SFPQ Gene Expression in Human induced Pluripotent Stem Cells Results in Massive Cell Death. Research Square. 1970 Jan 1;. Link: Research Square
  • Xiang, K & Bartel, D. (2021) The molecular basis of coupling between poly(A)-tail length and translational efficiency. bioRxiv. 1970 Jan 1;. Link: bioRxiv
  • Sugimoto, S, et al. (2021) An organoid-based organ-repurposing approach to treat short bowel syndrome. Nature. 1970 Jan 1;. PM ID: 33627870
  • Ma, P, et al. (2021) Avidity‐Based Selection of Tissue‐Specific CAR‐T Cells from a Combinatorial Cellular Library of CARs. Advanced Science. 1970 Jan 1;:2003091. Link: Advanced Science
  • Cho, JH, et al. (2021) Engineering advanced logic and distributed computing in human CAR immune cells. Nature communications. 1970 Jan 1; 12(1):792. PM ID: 33542232
  • Omori, S, et al. (2021) Tim4 recognizes carbon nanotubes and mediates phagocytosis leading to granuloma formation. Cell reports. 1970 Jan 1; 34(6):108734. PM ID: 33567275
  • Tjalsma, SJD, et al. (2021) H4K20me1 and H3K27me3 are concurrently loaded onto the inactive X chromosome but dispensable for inducing gene silencing. EMBO reports. 1970 Jan 1;:e51989. PM ID: 33605056
  • Kitano, H, et al. (2021) Development of a genetically modified hepatoma cell line with heat-inducible high liver function. Cytotechnology. 1970 Jan 1;. Link: Cytotechnology
  • Zhu, R, et al. (2021) Synthetic multistability in mammalian cells. bioRxiv. 1970 Jan 1;. Link: bioRxiv
  • Karlsson, J, et al. (2021) Photocrosslinked Bioreducible Polymeric Nanoparticles for Enhanced Systemic siRNA Delivery as Cancer Therapy. Advanced Functional Materials. 1970 Jan 1;:2009768. Link: Advanced Functional Materials