pMC.CMV-MCS-EF1α-GFP-SV40polyA Parental Minicircle Cloning Vector
- 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 | |||
|---|---|---|---|---|---|---|---|---|
| MN511A-1 | pMC.CMV-MCS-EF1α-GFP-SV40PolyA | 10 µg | $764 |
|
||||
Overview
Overview
Maximize safety with Minicircle Technology—episomal expression free from foreign DNA Get all the advantages of Minicircle Technology with the pMC.CMV-MCS-EF1α-GFP-SV40polyA Parental Minicircle Cloning Vector (Cat.# MN511A-1). This Minicircle Cloning Vector contains the necessary sequences to generate a minicircle, and a strong CMV promoter upstream of an MCS, so you can clone-in your gene-of-interest and get high expression levels in most commonly-used cell lines (i.e. HeLa, HEK293, HT1080). In addition, this vector includes GFP driven by the moderate EF1α promoter to facilitate identification of transfectants via GFP imaging.
About 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
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.
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
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.
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.
Resources
Citations
-
Johnston, CD, et al. (2019) Systematic evasion of the restriction-modification barrier in bacteria. Proc. Natl. Acad. Sci. U.S.A.. 2019 May 16;. PM ID: 31097593
-
Han, D, et al. (2019) Activation of Melatonin Receptor 2 But Not Melatonin Receptor 1 Mediates Melatonin-conferred Cardio-protection Against Myocardial Ischemia/Reperfusion Injury. J. Pineal Res.. 2019 Mar 22;:e12571. PM ID: 30903623
-
Petrini, S, et al. (2017) Aged induced pluripotent stem cell (iPSCs) as a new cellular model for studying premature aging. Aging (Albany NY). 2017 May 31; 9(5):1453-1469. PM ID: 28562315
-
Kelton, W, et al. (2017) Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange. Sci Rep. 2017 Apr 4; 7:45775. PM ID: 28374766
-
Traub, S, et al. (2017) Pharmaceutical Characterization of Tropomyosin Receptor Kinase B-Agonistic Antibodies on Human Induced Pluripotent Stem (hiPS) Cell-Derived Neurons. J. Pharmacol. Exp. Ther.. 2017 Jun 1; 361(3):355-365. PM ID: 28351853
-
Liu, N, et al. (2017) PIM1-minicircle as a therapeutic treatment for myocardial infarction. PLoS ONE. 2017 Mar 21; 12(3):e0173963. PM ID: 28323876
-
Zhang, Z, et al. (2017) Gene delivery of TIPE2 inhibits breast cancer development and metastasis via CD8(+) T and NK cell-mediated antitumor responses.. Mol. Immunol.. 2017 May 1; 85:230-237. PM ID: 28314212
-
Henno, L, et al. (2017) Analysis of Human Papillomavirus Genome Replication Using Two- and Three-Dimensional Agarose Gel Electrophoresis. Curr Protoc Microbiol. 2017 May 16; 45:14B.10.1-14B.10.37. PM ID: 28510360
-
Jaafar, L, et al. (2017) SFPQ•NONO and XLF function separately and together to promote DNA double-strand break repair via canonical nonhomologous end joining. Nucleic Acids Res.. 2017 Feb 28; 45(4):1848-1859. PM ID: 27924002
-
Wu, H, et al. (2017) MicroRNA-206 prevents hepatosteatosis and hyperglycemia by facilitating insulin signaling and impairing lipogenesis. J. Hepatol.. 2017 Apr 1; 66(4):816-824. PM ID: 28025059
-
Tidd, N, et al. (2017) Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells. Int J Mol Sci. 2017 Apr 12; 18(4). PM ID: 28417917
-
Brett, E, et al. (2017) Magnetic Nanoparticle-Based Upregulation of B-Cell Lymphoma 2 Enhances Bone Regeneration. Stem Cells Transl Med. 2017 Jan 1; 6(1):151-160. PM ID: 28170185
-
Tockner, B, et al. (2016) Construction and validation of an RNA trans-splicing molecule suitable to repair a large number of COL7A1 mutations. Gene Ther.. 2016 Nov 1; 23(11):775-784. PM ID: 27434145
-
Sun, JG, et al. (2016) Yap1 promotes the survival and self-renewal of breast tumor initiating cells via inhibiting Smad3 signaling. Oncotarget. 2016 Mar 1; 7(9):9692-706. PM ID: 26695440
-
Sharma, VS. (2016) Size controlled retinal differentiation of human induced pluripotent stem cells in shaking microwells. Thesis. ;. Link: Thesis
-
Dincer, E, et al. (2016) Canine Infections and Partial S Segment Sequence Analysis of Toscana Virus in Turkey. Vector Borne Zoonotic Dis.. 2016 Sep 1; 16(9):611-8. PM ID: 27400226
-
Gaspar, VM, et al. (2016) Highly selective capture of minicircle DNA biopharmaceuticals by a novel zinc-histidine peptide conjugate. Separation and Purification Technology. 2016 Oct 31; 174:417–424. Link: Separation and Purification Technology
-
Mofid, A. (2016) Ultrasound-Mediated S100A6 Gene Therapy Ameliorates Myocardial Ischemia/Reperfusion (I/R) Injury. Thesis. ;. Link: Thesis
-
Fernandes, AR & Chari, DM. (2016) Part II: Functional delivery of a neurotherapeutic gene to neural stem cells using minicircle DNA and nanoparticles: Translational advantages for regenerative neurology. J Control Release. 2016 Sep 28; 238:300-10. PM ID: 27369863
-
Fernandes, AR & Chari, DM. (2016) Part I: Minicircle vector technology limits DNA size restrictions on ex vivo gene delivery using nanoparticle vectors: Overcoming a translational barrier in neural stem cell therapy. J Control Release. 2016 Sep 28; 238:289-99. PM ID: 27317366
- See More
Related Products
Products
| Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
|---|---|---|---|---|---|---|---|---|
| MN511A-1 | pMC.CMV-MCS-EF1α-GFP-SV40PolyA | 10 µg | $764 |
|
||||
Overview
Overview
Maximize safety with Minicircle Technology—episomal expression free from foreign DNA Get all the advantages of Minicircle Technology with the pMC.CMV-MCS-EF1α-GFP-SV40polyA Parental Minicircle Cloning Vector (Cat.# MN511A-1). This Minicircle Cloning Vector contains the necessary sequences to generate a minicircle, and a strong CMV promoter upstream of an MCS, so you can clone-in your gene-of-interest and get high expression levels in most commonly-used cell lines (i.e. HeLa, HEK293, HT1080). In addition, this vector includes GFP driven by the moderate EF1α promoter to facilitate identification of transfectants via GFP imaging.About 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
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.
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
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.
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.
Citations
-
Johnston, CD, et al. (2019) Systematic evasion of the restriction-modification barrier in bacteria. Proc. Natl. Acad. Sci. U.S.A.. 2019 May 16;. PM ID: 31097593
-
Han, D, et al. (2019) Activation of Melatonin Receptor 2 But Not Melatonin Receptor 1 Mediates Melatonin-conferred Cardio-protection Against Myocardial Ischemia/Reperfusion Injury. J. Pineal Res.. 2019 Mar 22;:e12571. PM ID: 30903623
-
Petrini, S, et al. (2017) Aged induced pluripotent stem cell (iPSCs) as a new cellular model for studying premature aging. Aging (Albany NY). 2017 May 31; 9(5):1453-1469. PM ID: 28562315
-
Kelton, W, et al. (2017) Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange. Sci Rep. 2017 Apr 4; 7:45775. PM ID: 28374766
-
Traub, S, et al. (2017) Pharmaceutical Characterization of Tropomyosin Receptor Kinase B-Agonistic Antibodies on Human Induced Pluripotent Stem (hiPS) Cell-Derived Neurons. J. Pharmacol. Exp. Ther.. 2017 Jun 1; 361(3):355-365. PM ID: 28351853
-
Liu, N, et al. (2017) PIM1-minicircle as a therapeutic treatment for myocardial infarction. PLoS ONE. 2017 Mar 21; 12(3):e0173963. PM ID: 28323876
-
Zhang, Z, et al. (2017) Gene delivery of TIPE2 inhibits breast cancer development and metastasis via CD8(+) T and NK cell-mediated antitumor responses.. Mol. Immunol.. 2017 May 1; 85:230-237. PM ID: 28314212
-
Henno, L, et al. (2017) Analysis of Human Papillomavirus Genome Replication Using Two- and Three-Dimensional Agarose Gel Electrophoresis. Curr Protoc Microbiol. 2017 May 16; 45:14B.10.1-14B.10.37. PM ID: 28510360
-
Jaafar, L, et al. (2017) SFPQ•NONO and XLF function separately and together to promote DNA double-strand break repair via canonical nonhomologous end joining. Nucleic Acids Res.. 2017 Feb 28; 45(4):1848-1859. PM ID: 27924002
-
Wu, H, et al. (2017) MicroRNA-206 prevents hepatosteatosis and hyperglycemia by facilitating insulin signaling and impairing lipogenesis. J. Hepatol.. 2017 Apr 1; 66(4):816-824. PM ID: 28025059
-
Tidd, N, et al. (2017) Minicircle Mediated Gene Delivery to Canine and Equine Mesenchymal Stem Cells. Int J Mol Sci. 2017 Apr 12; 18(4). PM ID: 28417917
-
Brett, E, et al. (2017) Magnetic Nanoparticle-Based Upregulation of B-Cell Lymphoma 2 Enhances Bone Regeneration. Stem Cells Transl Med. 2017 Jan 1; 6(1):151-160. PM ID: 28170185
-
Tockner, B, et al. (2016) Construction and validation of an RNA trans-splicing molecule suitable to repair a large number of COL7A1 mutations. Gene Ther.. 2016 Nov 1; 23(11):775-784. PM ID: 27434145
-
Sun, JG, et al. (2016) Yap1 promotes the survival and self-renewal of breast tumor initiating cells via inhibiting Smad3 signaling. Oncotarget. 2016 Mar 1; 7(9):9692-706. PM ID: 26695440
-
Sharma, VS. (2016) Size controlled retinal differentiation of human induced pluripotent stem cells in shaking microwells. Thesis. ;. Link: Thesis
-
Dincer, E, et al. (2016) Canine Infections and Partial S Segment Sequence Analysis of Toscana Virus in Turkey. Vector Borne Zoonotic Dis.. 2016 Sep 1; 16(9):611-8. PM ID: 27400226
-
Gaspar, VM, et al. (2016) Highly selective capture of minicircle DNA biopharmaceuticals by a novel zinc-histidine peptide conjugate. Separation and Purification Technology. 2016 Oct 31; 174:417–424. Link: Separation and Purification Technology
-
Mofid, A. (2016) Ultrasound-Mediated S100A6 Gene Therapy Ameliorates Myocardial Ischemia/Reperfusion (I/R) Injury. Thesis. ;. Link: Thesis
-
Fernandes, AR & Chari, DM. (2016) Part II: Functional delivery of a neurotherapeutic gene to neural stem cells using minicircle DNA and nanoparticles: Translational advantages for regenerative neurology. J Control Release. 2016 Sep 28; 238:300-10. PM ID: 27369863
-
Fernandes, AR & Chari, DM. (2016) Part I: Minicircle vector technology limits DNA size restrictions on ex vivo gene delivery using nanoparticle vectors: Overcoming a translational barrier in neural stem cell therapy. J Control Release. 2016 Sep 28; 238:289-99. PM ID: 27317366
- See More