pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning Vector

Take advantage of minicircle technology when you deliver shRNA with this vector – drive your shRNA using the strong H1 promoter and select using GFP and puro
  • Episomal expression sustained over weeks
  • Foreign DNA-free
  • More efficient transfections from small plasmid size
  • Unlimited insert size
  • Optimized coli minicircle production strain

Products

Catalog Number Description Size Price Quantity Add to Cart
MNSI505A-1 pMC.CMV-GFP-T2A-Puro-H1-MCS (shRNA expression parental minicircle plasmid) 10 µg $808
- +

Overview

Overview

Get episomal expression of your shRNA without introducing foreign DNA to cells

Deliver shRNAs to cells with minicircles produced from the pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning Vector and get all the advantages of Minicircle Technology. With your shRNA driven by the strong H1 shRNA promoter and both GFP and puromycin resistance for selection of transfectants, you can deliver your shRNA-of-interest episomally on an efficiently-transfected plasmid that is free of foreign DNA.

The pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning Vector co-expresses the GFP and puromycin resistance selection markers via the strong CMV promoter.

pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning VectorAbout Minicircle Technology
  • Episomal expression sustained over weeks
  • Foreign DNA-free
  • More efficient transfections from small plasmid size
  • Unlimited insert size
  • Optimized coli minicircle production strain
  • Works in vitro and in vivo
When you want sustained transgene expression without introducing any foreign DNA—such as for model animal and gene therapy development—Minicircle Technology is a great gene expression option. Produced as small excised, circular DNA fragments from a parental plasmid, the non-viral, episomal Minicircle expression cassette is free of any bacterial plasmid DNA sequences, and comes with a variety of promoter and reporter combinations. Their small size facilitates more efficient transfection than what’s possible with standard-sized plasmids, and, while Minicircles do not replicate with the host cell, expression lasts for 14 days or longer in dividing cells, and can continue for months in non-dividing cells. Product Note: Parental minicircle plasmids and the ZYCY10P3S2T Producer Bacterial Strain are available for purchase by not-for-profit researchers only. Commercial users may purchase pre-made, ready-to-transfect minicircle DNA only. SBI also offers custom parental plasmid cloning and minicircle DNA production to both not-for-profit and commercial end users—contact services@systembio.com for additional details. For any other purposes, including the ability to buy the parental MC production system, commercial users should contact SBI at tech@systembio.com for further information.

How It Works

How It Works

Generating minicircles from the parental cloning vector

To generate minicircles that are ready for transfection, you need your Minicircle Cloning Vector with your insert (gene, promoter-gene cassette, small RNA, etc.), SBI’s optimized, ready-to-transform ZYCY10P3S2T E. coli Minicircle Producer Strain (Cat.# MN900A-1), and arabinose (Cat.# MN850A-1).

Minicircles are produced from the full-sized Parental Minicircle using PhiC31 Integrase, which mediates a recombination event between the PhiC321 attB and attP sites on the parental plasmid (Figure 1). This reaction results in two products—the minicircle, which is now free from any bacterial DNA sequences—and the parental plasmid. To get rid of the parental plasmid, the I-SceI endonuclease recognizes and acts on the I-SceI sites on the parental plasmid, resulting in degradation of the parental plasmid.

How to generate minicircles from the parental minicircle vector

Figure 1. Generating minicircle DNA from the Parental Minicircle Plasmid.

More about the ZYCY10P3S2T E. coli Minicircle Producer Strain

The Minicircle Producer Strain harbors an arabinose-inducible system to express the PhiC31 integrase and the I-SceI endonuclease simultaneously. The ZYCY10P3S2T strain also contains a robust arabinose transporter LacY A177C gene. Adding arabinose to the media turns on expression of the PhiC31 integrase and endonuclease genes, resulting in separation of the Parental Minicircle Plasmid into the individual minicircle and parental plasmids (from the PhiC31 Integrase activity), and the degradation of the parental plasmid (from Sce-1 endonuclease activity).

Supporting Data

Supporting Data

Achieve sustained expression from minicircles after transfection in vitro and in vivo

Use minicircles to get easy, sustained transfection in most cell types

Figure 1. Easy, sustained transfection in most cell types. Transfection of 1 μg of minicircle DNA (pMC.CMV-MCS-EF1-GFPSV40PolyA, Cat.# MN511A-1) into HEK293 cells delivers over one week of robust gene expression.

Use minicircles to express transgenes for weeks in animal models

Figure 2. Express transgenes for weeks in animal models. (A) Hydrodynamic tail vein injection of 2 µg and 4 µg of minicircle DNA (CMV-GFP-Luc) into mice shows excellent expression after 48 hours. (B) Minicircle-delivered transgenes retain robust expression that can last for weeks compared to transgenes that are delivered using plasmid DNA, where expression is rapidly lost. In this study, 40 µg of minicircle DNA was introduced into mice via hydrodynamic tail vein injection.

FAQs

Resources

Citations

  • Wang, H, et al. (2024) TMIGD2 is an orchestrator and therapeutic target on human acute myeloid leukemia stem cells. Nature communications. 2024; 15(1):11. PM ID: 38167704
  • Jia, W, et al. (2024) Microcystin-RR promote lipid accumulation through CD36 mediated signal pathway and fatty acid uptake in HepG2 cells. Environmental Research. 2024;:118402. Link: Environmental Research
  • O'Dwyer, M, et al. (2024) PSGL-1 decorated with sialyl Lewisa/x promotes high affinity binding of myeloma cells to P-selectin but is dispensable for E-selectin engagement. Scientific reports. 2024; 14(1):1756. PM ID: 38243063
  • Deng, R, et al. (2024) PZR suppresses innate immune response to RNA viral infection by inhibiting MAVS activation in interferon signaling mediated by RIG-I and MDA5. Antiviral research. 2024; 222:105797. PM ID: 38185222
  • Yan, C, et al. (2024) PD-L1 Expression Is Increased in LPS-Induced Acute Respiratory Distress Syndrome by PI3K-AKT-Egr-1/C/EBPδ Signaling Pathway. Inflammation. 2024;. PM ID: 38376609
  • Zeng, ZC, et al. (2023) METTL3 protects METTL14 from STUB1-mediated degradation to maintain m6 A homeostasis. EMBO reports. 2023;:e55762. PM ID: 36597993
  • Sierra-Magro, A, et al. (2023) C/EBPβ Regulates TFAM Expression, Mitochondrial Function and Autophagy in Cellular Models of Parkinson’s Disease. International journal of molecular sciences. 2023; 24(2). PM ID: 36674978
  • Ma, J, et al. (2023) Enhanced E6AP-mediated ubiquitination of ENO1 via LINC00663 contributes to radiosensitivity of breast cancer by regulating mitochondrial homeostasis. Cancer Letters. 2023;:216118. Link: Cancer Letters
  • Wen, J, et al. (2023) Lnc-17Rik promotes the immunosuppressive function of Myeloid-Derived suppressive cells in esophageal cancer. Cellular immunology. 2023; 385:104676. PM ID: 36780770
  • Hasegawa, S, Imai, M & Takahashi, N. (2023) Role of acetoacetyl-CoA synthetase in glucose uptake by HepG2 cells. bioRxiv. 2023;. Link: bioRxiv
  • Agarwal, S, et al. (2023) BZW2 Inhibition Reduces Colorectal Cancer Growth and Metastasis. Molecular cancer research : MCR. 2023;:OF1-OF15. PM ID: 37067340
  • Song, S, et al. (2023) CHMP4A stimulates CD8+ T-lymphocyte infiltration and inhibits breast tumor growth via the LSD1/IFNβ axis. Cancer science. 2023;. PM ID: 37198999
  • Shaker, BT, et al. (2023) The 14-Kilodalton Human Growth Hormone Fragment a Potent Inhibitor of Angiogenesis and Tumor Metastasis. International journal of molecular sciences. 2023; 24(10). PM ID: 37240223
  • Ju, Z, et al. (2023) TXNL4B regulates radioresistance by controlling the PRP3-mediated alternative splicing of FANCI. MedComm. 2023; 4(3):e258. PM ID: 37168687
  • Salehi, S, et al. (2023) Cytosolic Ptbp2 modulates axon growth in motoneurons through axonal localization and translation of Hnrnpr. Nature communications. 2023; 14(1):4158. PM ID: 37438340
  • Zhang, B, et al. (2023) WIF1 promoter hypermethylation induce endometrial carcinogenesis through the Wnt/β-catenin signaling pathway. American journal of reproductive immunology (New York, N.Y. : 1989). 2023; 90(2):e13743. PM ID: 37491917
  • Xu, W, et al. (2023) Exosomal PIK3CB promotes PD-L1 expression and malignant transformation in esophageal squamous cell carcinoma. Medical oncology (Northwood, London, England). 2023; 40(8):221. PM ID: 37402056
  • GUAN, Y, et al. (2023) Ginsenoside Rg1 protects against ischemia-induced neuron damage by regulating the rno-miRNA-27a-3p/PPARγ axis. BIOCELL. 2023; 47(7):1583-1594. Link: BIOCELL
  • Maruyama, M, et al. (2023) Neat1 lncRNA organizes the inflammatory gene expressions in the dorsal root ganglion in neuropathic pain caused by nerve injury. Frontiers in immunology. 2023; 14:1185322. PM ID: 37614230
  • Ma, Y, et al. (2023) The DACH1 Gene Transcriptional Activation and Protein Degradation Mediated by Transactivator Tas of Prototype Foamy Virus. Viruses. 2023; 15(9). PM ID: 37766305
pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning Vector $808.00

Products

Catalog Number Description Size Price Quantity Add to Cart
MNSI505A-1 pMC.CMV-GFP-T2A-Puro-H1-MCS (shRNA expression parental minicircle plasmid) 10 µg $808
- +

Overview

Overview

Get episomal expression of your shRNA without introducing foreign DNA to cells

Deliver shRNAs to cells with minicircles produced from the pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning Vector and get all the advantages of Minicircle Technology. With your shRNA driven by the strong H1 shRNA promoter and both GFP and puromycin resistance for selection of transfectants, you can deliver your shRNA-of-interest episomally on an efficiently-transfected plasmid that is free of foreign DNA.

The pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning Vector co-expresses the GFP and puromycin resistance selection markers via the strong CMV promoter.

pMC.CMV-GFP-T2A-Puro-H1-MCS Parental Minicircle shRNA Cloning VectorAbout Minicircle Technology
  • Episomal expression sustained over weeks
  • Foreign DNA-free
  • More efficient transfections from small plasmid size
  • Unlimited insert size
  • Optimized coli minicircle production strain
  • Works in vitro and in vivo
When you want sustained transgene expression without introducing any foreign DNA—such as for model animal and gene therapy development—Minicircle Technology is a great gene expression option. Produced as small excised, circular DNA fragments from a parental plasmid, the non-viral, episomal Minicircle expression cassette is free of any bacterial plasmid DNA sequences, and comes with a variety of promoter and reporter combinations. Their small size facilitates more efficient transfection than what’s possible with standard-sized plasmids, and, while Minicircles do not replicate with the host cell, expression lasts for 14 days or longer in dividing cells, and can continue for months in non-dividing cells. Product Note: Parental minicircle plasmids and the ZYCY10P3S2T Producer Bacterial Strain are available for purchase by not-for-profit researchers only. Commercial users may purchase pre-made, ready-to-transfect minicircle DNA only. SBI also offers custom parental plasmid cloning and minicircle DNA production to both not-for-profit and commercial end users—contact services@systembio.com for additional details. For any other purposes, including the ability to buy the parental MC production system, commercial users should contact SBI at tech@systembio.com for further information.

How It Works

How It Works

Generating minicircles from the parental cloning vector

To generate minicircles that are ready for transfection, you need your Minicircle Cloning Vector with your insert (gene, promoter-gene cassette, small RNA, etc.), SBI’s optimized, ready-to-transform ZYCY10P3S2T E. coli Minicircle Producer Strain (Cat.# MN900A-1), and arabinose (Cat.# MN850A-1).

Minicircles are produced from the full-sized Parental Minicircle using PhiC31 Integrase, which mediates a recombination event between the PhiC321 attB and attP sites on the parental plasmid (Figure 1). This reaction results in two products—the minicircle, which is now free from any bacterial DNA sequences—and the parental plasmid. To get rid of the parental plasmid, the I-SceI endonuclease recognizes and acts on the I-SceI sites on the parental plasmid, resulting in degradation of the parental plasmid.

How to generate minicircles from the parental minicircle vector

Figure 1. Generating minicircle DNA from the Parental Minicircle Plasmid.

More about the ZYCY10P3S2T E. coli Minicircle Producer Strain

The Minicircle Producer Strain harbors an arabinose-inducible system to express the PhiC31 integrase and the I-SceI endonuclease simultaneously. The ZYCY10P3S2T strain also contains a robust arabinose transporter LacY A177C gene. Adding arabinose to the media turns on expression of the PhiC31 integrase and endonuclease genes, resulting in separation of the Parental Minicircle Plasmid into the individual minicircle and parental plasmids (from the PhiC31 Integrase activity), and the degradation of the parental plasmid (from Sce-1 endonuclease activity).

Supporting Data

Supporting Data

Achieve sustained expression from minicircles after transfection in vitro and in vivo

Use minicircles to get easy, sustained transfection in most cell types

Figure 1. Easy, sustained transfection in most cell types. Transfection of 1 μg of minicircle DNA (pMC.CMV-MCS-EF1-GFPSV40PolyA, Cat.# MN511A-1) into HEK293 cells delivers over one week of robust gene expression.

Use minicircles to express transgenes for weeks in animal models

Figure 2. Express transgenes for weeks in animal models. (A) Hydrodynamic tail vein injection of 2 µg and 4 µg of minicircle DNA (CMV-GFP-Luc) into mice shows excellent expression after 48 hours. (B) Minicircle-delivered transgenes retain robust expression that can last for weeks compared to transgenes that are delivered using plasmid DNA, where expression is rapidly lost. In this study, 40 µg of minicircle DNA was introduced into mice via hydrodynamic tail vein injection.

FAQs

Citations

  • Wang, H, et al. (2024) TMIGD2 is an orchestrator and therapeutic target on human acute myeloid leukemia stem cells. Nature communications. 2024; 15(1):11. PM ID: 38167704
  • Jia, W, et al. (2024) Microcystin-RR promote lipid accumulation through CD36 mediated signal pathway and fatty acid uptake in HepG2 cells. Environmental Research. 2024;:118402. Link: Environmental Research
  • O'Dwyer, M, et al. (2024) PSGL-1 decorated with sialyl Lewisa/x promotes high affinity binding of myeloma cells to P-selectin but is dispensable for E-selectin engagement. Scientific reports. 2024; 14(1):1756. PM ID: 38243063
  • Deng, R, et al. (2024) PZR suppresses innate immune response to RNA viral infection by inhibiting MAVS activation in interferon signaling mediated by RIG-I and MDA5. Antiviral research. 2024; 222:105797. PM ID: 38185222
  • Yan, C, et al. (2024) PD-L1 Expression Is Increased in LPS-Induced Acute Respiratory Distress Syndrome by PI3K-AKT-Egr-1/C/EBPδ Signaling Pathway. Inflammation. 2024;. PM ID: 38376609
  • Zeng, ZC, et al. (2023) METTL3 protects METTL14 from STUB1-mediated degradation to maintain m6 A homeostasis. EMBO reports. 2023;:e55762. PM ID: 36597993
  • Sierra-Magro, A, et al. (2023) C/EBPβ Regulates TFAM Expression, Mitochondrial Function and Autophagy in Cellular Models of Parkinson’s Disease. International journal of molecular sciences. 2023; 24(2). PM ID: 36674978
  • Ma, J, et al. (2023) Enhanced E6AP-mediated ubiquitination of ENO1 via LINC00663 contributes to radiosensitivity of breast cancer by regulating mitochondrial homeostasis. Cancer Letters. 2023;:216118. Link: Cancer Letters
  • Wen, J, et al. (2023) Lnc-17Rik promotes the immunosuppressive function of Myeloid-Derived suppressive cells in esophageal cancer. Cellular immunology. 2023; 385:104676. PM ID: 36780770
  • Hasegawa, S, Imai, M & Takahashi, N. (2023) Role of acetoacetyl-CoA synthetase in glucose uptake by HepG2 cells. bioRxiv. 2023;. Link: bioRxiv
  • Agarwal, S, et al. (2023) BZW2 Inhibition Reduces Colorectal Cancer Growth and Metastasis. Molecular cancer research : MCR. 2023;:OF1-OF15. PM ID: 37067340
  • Song, S, et al. (2023) CHMP4A stimulates CD8+ T-lymphocyte infiltration and inhibits breast tumor growth via the LSD1/IFNβ axis. Cancer science. 2023;. PM ID: 37198999
  • Shaker, BT, et al. (2023) The 14-Kilodalton Human Growth Hormone Fragment a Potent Inhibitor of Angiogenesis and Tumor Metastasis. International journal of molecular sciences. 2023; 24(10). PM ID: 37240223
  • Ju, Z, et al. (2023) TXNL4B regulates radioresistance by controlling the PRP3-mediated alternative splicing of FANCI. MedComm. 2023; 4(3):e258. PM ID: 37168687
  • Salehi, S, et al. (2023) Cytosolic Ptbp2 modulates axon growth in motoneurons through axonal localization and translation of Hnrnpr. Nature communications. 2023; 14(1):4158. PM ID: 37438340
  • Zhang, B, et al. (2023) WIF1 promoter hypermethylation induce endometrial carcinogenesis through the Wnt/β-catenin signaling pathway. American journal of reproductive immunology (New York, N.Y. : 1989). 2023; 90(2):e13743. PM ID: 37491917
  • Xu, W, et al. (2023) Exosomal PIK3CB promotes PD-L1 expression and malignant transformation in esophageal squamous cell carcinoma. Medical oncology (Northwood, London, England). 2023; 40(8):221. PM ID: 37402056
  • GUAN, Y, et al. (2023) Ginsenoside Rg1 protects against ischemia-induced neuron damage by regulating the rno-miRNA-27a-3p/PPARγ axis. BIOCELL. 2023; 47(7):1583-1594. Link: BIOCELL
  • Maruyama, M, et al. (2023) Neat1 lncRNA organizes the inflammatory gene expressions in the dorsal root ganglion in neuropathic pain caused by nerve injury. Frontiers in immunology. 2023; 14:1185322. PM ID: 37614230
  • Ma, Y, et al. (2023) The DACH1 Gene Transcriptional Activation and Protein Degradation Mediated by Transactivator Tas of Prototype Foamy Virus. Viruses. 2023; 15(9). PM ID: 37766305