pMC.BESPX-MCS2 Empty 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 | |||
---|---|---|---|---|---|---|---|---|
MN100B-1 | pMC.BESPX-MCS2 (empty vector) | 10 µg | $670 |
|
Overview
Overview
Maximize safety with Minicircle Technology—episomal expression free from foreign DNA Make the most of Minicircle Technology with the pMC.BESPX-MCS2 Empty Parental Minicircle Cloning Vector (Cat.# MN100B-1. This Minicircle Cloning Vector contains only the necessary sequences to generate a minicircle and an MCS—MCS2—so you can clone in your own promoter-gene expression cassette.
- 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.
FAQs
Resources
Related Products
Citations
-
Johnston, CD, et al. (2019) Systematic evasion of the restriction-modification barrier in bacteria. Proc. Natl. Acad. Sci. U.S.A.. 2019;. 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;: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; 9(5):1453-1469. PM ID: 28562315
-
Kelton, W, et al. (2017) Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange. Sci Rep. 2017; 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; 361(3):355-365. PM ID: 28351853
-
Liu, N, et al. (2017) PIM1-minicircle as a therapeutic treatment for myocardial infarction. PLoS ONE. 2017; 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; 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; 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; 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; 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; 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; 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; 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; 7(9):9692-706. PM ID: 26695440
-
Sharma, VS. (2016) Size controlled retinal differentiation of human induced pluripotent stem cells in shaking microwells. Thesis. 2016;. 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; 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; 174:417–424. Link: Separation and Purification Technology
-
Mofid, A. (2016) Ultrasound-Mediated S100A6 Gene Therapy Ameliorates Myocardial Ischemia/Reperfusion (I/R) Injury. Thesis. 2016;. 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; 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; 238:289-99. PM ID: 27317366
- See More
Products
Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
---|---|---|---|---|---|---|---|---|
MN100B-1 | pMC.BESPX-MCS2 (empty vector) | 10 µg | $670 |
|
Overview
Overview
Maximize safety with Minicircle Technology—episomal expression free from foreign DNA Make the most of Minicircle Technology with the pMC.BESPX-MCS2 Empty Parental Minicircle Cloning Vector (Cat.# MN100B-1. This Minicircle Cloning Vector contains only the necessary sequences to generate a minicircle and an MCS—MCS2—so you can clone in your own promoter-gene expression cassette.
- 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.
FAQs
Citations
-
Johnston, CD, et al. (2019) Systematic evasion of the restriction-modification barrier in bacteria. Proc. Natl. Acad. Sci. U.S.A.. 2019;. 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;: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; 9(5):1453-1469. PM ID: 28562315
-
Kelton, W, et al. (2017) Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange. Sci Rep. 2017; 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; 361(3):355-365. PM ID: 28351853
-
Liu, N, et al. (2017) PIM1-minicircle as a therapeutic treatment for myocardial infarction. PLoS ONE. 2017; 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; 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; 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; 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; 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; 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; 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; 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; 7(9):9692-706. PM ID: 26695440
-
Sharma, VS. (2016) Size controlled retinal differentiation of human induced pluripotent stem cells in shaking microwells. Thesis. 2016;. 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; 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; 174:417–424. Link: Separation and Purification Technology
-
Mofid, A. (2016) Ultrasound-Mediated S100A6 Gene Therapy Ameliorates Myocardial Ischemia/Reperfusion (I/R) Injury. Thesis. 2016;. 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; 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; 238:289-99. PM ID: 27317366
- See More