pMC.CMV-GFP-H1-MCS Parental Minicircle shRNA 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 | |||
|---|---|---|---|---|---|---|---|---|
| MNSI500A-1 | pMC.CMV-GFP-H1-MCS (shRNA expression parental minicircle plasmid) | 10 µg | $808 |
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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-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 GFP for selection of transfectants, you can deliver your shRNA-of-interest episomally on an efficiently-transfected plasmid that is free of foreign DNA.
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
References
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
Citations
Related Products
Products
| Catalog Number | Description | Size | Price | Quantity | Add to Cart | |||
|---|---|---|---|---|---|---|---|---|
| MNSI500A-1 | pMC.CMV-GFP-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-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 GFP for selection of transfectants, you can deliver your shRNA-of-interest episomally on an efficiently-transfected plasmid that is free of foreign DNA.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
References
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.