Minicircle Technology

Minicircle DNA Vector Technology Non-integrative sustained expression

The Minicircle Advantage Highlights

Minicircles are episomal DNA vectors that are produced as circular expression Expression for up to 14 days cassettes devoid of any bacterial DNA backbone. Their smaller molecular in dividing cells. Even longer size enables more efficient and offers sustained expression over a for non-dividing cells period of weeks as compared to standard plasmid vectors that only work for a few days. ZYCY10P3S2T E. coli cells available for minicircle Production of Minicircle DNA production For use in vivo or in vitro 1 Clone your gene-of-interest into the parental plasmid Choose your promoter, 2 Transform into ZYCY10P3S2T E. coli attR reporter gene, and vector format 3 Induce with arabinose Parental plasmid is SV40 degraded, preventing 4 Purify with plasmid purification kit poly-A Minicircle immune responses

attB Promoter pUC ORI 32x Sce-I Sites ZYCY10P3S2T E. coli Promoter Bacterial strain engineered for Parental + Arabinose minicircle production. KanR (switches on ΦC31 Integrase Arabinose induces ФC31 Plasmid and Sce-I Endonuclease genes) integrase and the I-SceI endonuclease simultaneously. ФC31 integrase produces the

Transgene MC-DNA molecules and the attP SV40 pUC ORI parental plasmid. poly-A 32x Sce-I Sites SceI endonuclease degrades attL Arabinose_ Bacterial the parental plasmid DNA M + Backbone backbone. 10kb PP As described in KanR 3kb MC Mark A. Kay, Cheng-Yi He & Zhi-Ying Chen. A robust system for production of minicircle DNA 1kb Degradation vectors. Nature Biotechnology, (2010). doi:10.1038/nbt.1708. www.systembio.com/minicircles Minicircle DNA Technology

Minicircle Vector Offering Expression cassette options SV40 Cat # poly-A CMV MCS MN501A-1 Only from attB SBI ! 32x Sce-I EF1a MCS MN502A-1 Sites E x p r CMV MCS EF1a GFP pUC ORI e MN511A-1

s

s

i

o

R Parental RFP

n CMV MCS EF1a MN512A-1

c

Kan

a Supporting Products Plasmid s

s GFP

e EF1a MCS IRES MN530A-1

t Kanamycin (50mg/ml) MN800A-1

t

e

o

p Arabinose solution (20%) MN850A-1

t EF1a MCS IRES RFP i MN531A-1

o n s positive control ZYCY10P3S2T E. coli MN900A-1 attP CMV GFP MN601A-1

positive control CMV RFP MN602A-1

Long-term in vitro expression from 6 hrs 2 days 8 days Minicircle Minicircle

Minicircle GFP GFP GFP Minicircle

GFP

Sustained in vivo expression 10,000 Minicircle DNA Plasmid DNA

1,000

100 Mouse Tail-vein injection leads to sustained

Transgene Expression Level Expression Transgene 1 48 Hours 2 Weeks 3 Weeks expression for months We Also Offer Custom Services System Biosciences offers a wide-range of custom services to support your research, allowing you to spend less time making tools, and more time making discoveries. To learn more, visit our website at www.systembio.com/service or call us at 888-266-5066.

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