OverviewSimplifying minicircle production Optimized for minicircle production, the ZYCY10P3S2T Minicircle Production Strain comes as competent cells, so you can directly transform ligation reactions or plasmids. The competent cells are prepared and tested using a modification of the procedure developed by Kay, et al.1, and are suitable for cloning and production of minicircle-based plasmids.The ZYCY10P3S2T E.coli Minicircle Production Strain is derived from the BW27783 E. coli strain that stably expresses a set of inducible minicircle-assembly enzymes, PhiC31 integrase, and I-SceI homing endonuclease. This bacterial strain enables production and purification of minicircles in a time frame and quantity similar to that of routine plasmid DNA preparation.Please note that other competent E. coli strains cannot be substituted for minicircle production as they do not have the appropriate genomic modifications to produce minicircles.About Minicircle TechnologyEpisomal expression sustained over weeksForeign DNA-freeMore efficient transfections from small plasmid sizeUnlimited insert sizeOptimized coli minicircle production strainWorks in vitro and in vivoWhen 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.ReferencesKay MA, He CY, and Chen ZY. A robust system for production of minicircle DNA vectors. Nat Biotechnol. 2010 Dec; 28(12):1287-9. PMCID: PMC4144359.How It WorksGenerating minicircles from the parental cloning vectorTo 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 StrainThe 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 I-SceI endonuclease activity).Supporting DataAchieve sustained expression from minicircles after transfection in vitro and in vivoFigure 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.ResourcesUser Manual: Minicircle DNA Vector TechnologyProduct Sheet: Minicircle TechnologyBrochure: Gene Delivery and Expression Products and ServicesCitationsJohnston, 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:31097593Han, 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:30903623Traub, 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:28351853Petrini, 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:28562315Henno, 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:28510360Zhang, 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:28314212Tidd, 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:28417917Kelton, W, et al. (2017) Reprogramming MHC specificity by CRISPR-Cas9-assisted cassette exchange. Sci Rep.2017 Apr 4; 7:45775. PM ID:28374766Wu, 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:28025059Liu, N, et al. (2017) PIM1-minicircle as a therapeutic treatment for myocardial infarction. PLoS ONE.2017 Mar 21; 12(3):e0173963. PM ID:28323876Jaafar, 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:27924002Brett, 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:28170185Colluru, VT, Zahm, CD & McNeel, DG. (2016) Mini-intronic plasmid vaccination elicits tolerant LAG3(+) CD8(+) T cells and inferior antitumor responses. Oncoimmunology.2016 Nov 17; 5(10):e1223002. PM ID:27853647Li, F, et al. (2016) Minicircle HBV cccDNA with a Gaussia luciferase reporter for investigating HBV cccDNA biology and developing cccDNA-targeting drugs. Sci Rep.2016 Nov 7; 6:36483. PM ID:27819342Tockner, 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:27434145Gaspar, 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 TechnologyFernandes, 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:27317366Fernandes, 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:27369863Mun, JY, et al. (2016) Minicircle microporation-based non-viral gene delivery improved the targeting of mesenchymal stem cells to an injury site.. Biomaterials.2016 Sep 1; 101:310-20. PM ID:27315214Diamantino, T, et al. (2016) Minicircle DNA purification using a CIM® DEAE-1 monolithic support. J Sep Sci.2016 Sep 1; 39(18):3544-9. 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美国SBI代理

System Biosciences,简称SBI，美国加州湾区新成立的生技公司，致力于独特、创新生物技术之开发，以研发利于基因及蛋白质功能鉴定、研究之崭新方法和工具为宗旨。 现阶段研发重心为RNA干扰（RNAi）研究之相关工具。 System Biosciences (SBI) 致力于开发独特、革新的技术，为客户研究蛋白组学和基因组学功能提供研究工具。SBI 是专业的慢病毒产品公司，提供基于慢病毒

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ExoQuick and ExoQuick-TC Products

Product	Size	Catalog #
ExoQuick serum exosome precipitation solution (5 ml)	75 reactions	EXOQ5A-1
ExoQuick Plasma prep and Exosome precipitation kit (5 ml ExoQuick plus 500 ul Thrombin at 500U/mL), replaces EXOQ5TD-1 product	75 reactions	EXOQ5TM-1
Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL), replaces TDEXO-1 product	100 reactions	TMEXO-1
ExoQuick serum exosome precipitation solution (20 ml)	300 reactions	EXOQ20A-1
ExoQuick-TC for Tissue Culture Media and Urine (10ml)	10 reactions	EXOTC10A-1
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Exosome-depleted FBS Media Supplement - USA Certified	50 ml	EXO-FBS-50A-1
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ExoQuick-LP Lipoprotein Pre-Clear & Exosome Precipitation Kit	5 reactions	EXOLP5A-1