Millipore Sigma Vibrant Logo

SCR724 Simplicon® Cloning Vector (E3L)

Simplicon® Cloning Vector (E3L) (Part #: SCR724) is a novel system offering immediate highly sustained and tunable in vivo protein expression of the target gene(s) in transfected cells without the risk of genome integration.

Simplicon® Cloning Vector (E3L): のSDS（安全データシート）、CoA（試験成績書）およびCoQ（品質証明書）、カタログなど製品関連の技術資料を入手いただけます。
View This Product on Sigma-Aldrich
SCR724
1 vial  
価格を検索中…
価格が見つかりません
Minimum Quantity needs to be mulitiple of
Maximum Quantity is
弊社照会 詳細を表示　
値引
()
 
—
弊社照会
—
現在国内在庫なし 現在国内在庫なし
現在国内在庫有り 
販売中止
在庫僅少
現在国内在庫あり
    Remaining : Will advise
      Remaining : Will advise
      注文対象外
      お問合せください
      Contact Customer Service

       

      お問合せください

      概要

      Replacement Information
      Description
      Catalogue NumberSCR724
      DescriptionSimplicon® Cloning Vector (E3L)
      Background InformationSimplicon® is a novel system to effect immediate high sustained protein expression of multiple genes into transfected cells without the risk of genome integration. The technology employs a single, synthetic, polycistronic, self-replicating RNA based on the Venezuelan equine encephalitis (VEE) genome1,2,3,4. The Simplicon® RNA contains only genes encoding the VEE RNA replication machinery while the structural proteins that are required to make an infectious particle have been removed and replaced with the transgenes of interest. The Simplicon® RNA is a synthetic RNA generated from the Simplicon® Cloning Vector (E3L) plasmid.

      Introduction and replication of the Simplicon® RNA is expected to elicit a strong interferon response in transfected cells. To suppress the IFN responses, a Vaccinia virus protein5, B18R, is used for the original Simplicon® technology. Recently, we found that another Vaccinia virus protein5, E3L, also suppresses the IFN responses in Simplicon® RNA expression. B18R neutralizes type I interferons by direct binding, while E3L inhibits the cytoplasmic signaling pathways of IFN responses. Therefore, B18R and E3L are both employed in the Simplicon® Expression System and work collaboratively to suppress IFN responses. As a result, there is increased cell viability during RNA transfection and increased expression of the transgenes. The Simplicon® Expression System works in human cells and is not expected to work in mouse cells. This is because the B18R does not effectively neutralize mouse interferon (IFN)-β.

      One day after transfection of the Simplicon® RNA, a spike in the levels of transgenes can be observed. The expression levels are maintained by addition of B18R, E3L and the selective agent, puromycin throughout the duration of the experiment. Over time, expression levels are expected to diminish and stabilize to 1/5 – 1/10 the levels initially observed and may be close to physiological levels after one week. Expression levels and duration may change depending upon the cell types, transgenes and media conditions used. The Simplicon® technology has been successfully utilized for efficient human iPSC generation through the sustained expression of critical reprogramming factors3,4 and in the creation of cell lines that express and retain the metabolic activities of five cytochrome P450 enzymes6.

      In the Simplicon® Expression System, B18R and E3L are provided as a B18R-E3L RNA (Cat. No. SCR722) for the suppression of IFN responses at RNA transfection. For sustained transgene expression, recombinant B18R protein (Cat. No. SCR156 and SCR197) or B18R conditioned medium (B18R-CM) can be used. E3L is continuously provided from the Simplicon® RNA itself. B18R-CM can be produced from B18R-E3L RNA or B18R RNA synthesized using the B18R-E3L plasmid (Cat. No. SCR727) or B18R plasmid (Cat. No. SCR728), respectively.

      The Simplicon® Cloning Vector (E3L) (Cat. No. SCR724) is designed for expression of the transgenes of interest as a Simplicon® RNA. The vector encodes four non-structural replication complex proteins (nsPs) as a single ORF at the 5’ half of the RNA, and multiple cloning site, E3L and puromycin resistance gene at the 3’ half of the RNA. Simplicon® Cloning Vector (E3L) can accommodate transgene cassettes up to a total size of 8,300 bp. The presence of E3L in the Simplicon® RNA works to suppress the IFN responses for sustained expression of the Simplicon® RNA. For suppression of IFN responses at RNA transfection, we recommend B18R-E3L RNA (SCR722) prepared from the B18R-E3L plasmid (SCR727).
      References
      Product Information
      Components
      • Simplicon® Cloning Vector (Cat. # SCR724). One (1) vial containing 10 µL of Plasmid (1 µg/µL). Store at -20 °C.
      Applications
      ApplicationSimplicon® Cloning Vector (E3L) (Part #: SCR724) is a novel system offering immediate highly sustained and tunable in vivo protein expression of the target gene(s) in transfected cells without the risk of genome integration.
      Biological Information
      ConcentrationPlease refer to datasheet.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsSimplicon® Cloning Vector (Cat. # SCR724). One (1) vial containing 10 µL of Plasmid (1 µg/µL). Store at -20 °C.
      Packaging Information
      Material Size1 vial
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      カタログ番号 GTIN
      SCR724 04054839501739

      Documentation

      Simplicon® Cloning Vector (E3L) (M)SDS

      タイトル

      英語版製品安全データシート（(M)SDS） 

      Simplicon® Cloning Vector (E3L) 試験成績書（CoA）

      タイトルロット番号
      Simplicon™ Cloning Vector (E3L) - 3394145 3394145
      Simplicon™ Cloning Vector (E3L) - 3485562 3485562

      データシート

      タイトル
      Simplicon¿ Cloning Vector (E3L)

      ユーザーガイド

      タイトル
      SimpliconTM Expression System: Designing Cloning and RNA Synthesis for Expression of Self-Replicative RNA

      ベクター配列

      タイトル
      Simplicon Cloning Vector (E3L)