• shRNA and Cre Recombinase Cloning Service


    Vigene provides customized services for the designing of constructs based on either existing Genebank gene sequences, or customer provided shRNA sequence.

    Vigene shRNA Advantages and Service Options

    shRNA vectors: Adenoviral, lentiviral and rAAV vector available; from $595

    shRNA fully customizable cloning options

    4-in-1 shRNA; from $795

    Cre Inducible shRNA expression; from $595

    GFP or RFP expression markers available.

    Guaranteed knock down expression by at least 70%

    Vigene provides a set four expression constructs against every target gene with the guarantee that at least one of the four will have a knockdown effect of 70% or more on corresponding gene expression, as determined by qRT-PCR, otherwise Vigene will replace the constructs one time free of charge.

    shRNA Vectors and Viral Packaging Services

    Vigene is specialized in AAV, adenovirus and lentivirus packaging service, ranging from small crude scale to the custom large purified scale.

    We like to work with customers to plan the viral delivery of shRNA. For most customers, we will ship 4 shRNAs in plasmid for you to test. After finding the best one suit your experiments, you can then ask us to package the best one into AAV, adenovirus or lentivirus.

    AAV shRNA Cloning Service

    To express shRNA with rAAV, Vigene provides the choices of either U6 or H1 promoter to drive the shRNA expression and GFP or RFP as mammalian expression marker. shRNA design for your gene of interest (GOI) is also available, please request a quote and describe your project and design requirements.

    shRNA Vector Services

    Vigene offers shRNA construct in either plasmid, adenoviral, AAV, lentiviral vector. Providing 4 constructs + free scrambled control, 5 µg DNA. Vigene can clone these into the viral vector of your choice. Explore viral vector options.

     

    Cat.# Description Price

    C100004

    shRNA construct in plasmid vector, 4 constructs per set, 5 µg DNA; delivery ~2 weeks

    $595

    C100005

    shRNA construct in adenoviral vector, 4 constructs per set, 5 µg DNA; delivery ~3 weeks

    $995

    C100006

    shRNA construct in lentiviral vector, 4 constructs per set, 5 µg DNA; delivery ~2 weeks

    $595

    C100007

    shRNA construct in AAV vector, 4 constructs per set, 5 µg DNA; delivery ~2 weeks

    $595

     

    shRNA 4-in-1 Cloning Service

    Tired of testing 4 shRNAs? Vigene's proprietary technology puts
    4 shRNAs in one expression vector, + free scrambled control.
    Multiple shRNAs expressed from multiple shRNA expression
    cassettes within one vector has been proven to be an
    effective method in gene knock down, both in vitro or in vivo.
    This approach shows great advantage when several genes
    need to be suppressed at the same time.

    shRNA_image2

    Key Advantages of the 4in1 shRNA system

    ● Knock down up to 4 different genes using one single plasmid vector.

    ● 4in1 design is more effective than conventional single shRNA designs.

    ● Guaranteed to knock down expression of your gene by at least 70%.

    ● Only need to package one virus instead of multiple viruses, saving you cost and time.

     

    Cat.# Description Price

    C100008

    4in1 shRNA: 4 shRNA sequences in pAV-4in1-shRNA-GFP, 5 µg DNA delivery ~2-3 weeks

    $795

    4in1 shRNA Custom Service

    We provide the most customizable service. Our scientists will work with you step-by-step and learn about your research goal to design and construct the 4in1 shRNA vector in either the pAV-4in1shRNA-GFP.

    4in1 shRNA in the adenoviral or lentiviral system

    Vigene scientists can tailor your custom design to work with the adenoviral or lentiviral system (price on application). For more information or any questions, please contact us or request a quote.

    4in1 shRNA in action

    shRNA_4in1

    shRNAs (shRNA1-4) against NM_004414 showed repression of protein expression at various degree. When 4 shRNA expressed from one vector(4in1 shRNA), protein expression showed consistent and clear down regulation (shown in A.).

    4in1 shRNA down regulated GFP expression

    4in1 shRNA targeting GFP could completely block the GFP expression 48hrs after coexpression of GFP and 4in1shRNA in HEK293t cells, shown in B. top panel, while control 4in1 shNRA showed no effect on GFP expression in the lower panel.

     

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    Cre Dependent Services

    Vigene Biosciences provides multiple options for Cre dependent shRNA expression and viral vector systems.

    Withthe tight structure of RNA polymerase III promoter, it is difficult to build an inducible promoter system for conventional shRNA expression driven by U6 or H1 promoter. The development of the microRNA scaffold based shRNA expression system, RNA polymerase II promoters can then be used for shRNA expression.

    Tet-on/off promoter system, as one example, is leaky, and an extra tTA gene has to be introduced. The tet-on/off is not an ideal system especially for in vivo use. With the existence of many Cre mouse lines and Cre dependent gene expression, the FLEX system has proven to be very powerful in controling in vivo gene expression.

    There are very few reports on using the FLEX system in controling Cre dependent shRNA expression. One of the reasons is due to the hairpin structure, sense and antisense strand of RNA can both produce the functional shRNA. This makes the FLEX system useless. Using Vigene's system, which is base on a modified FLEX system and microRNA shRNA scaffold, this can be overcome.

    Expression systems

    Conventional and inducible shRNA expression systems

    Cre Dependent shRNA expression system

    pAV-CreOn-shRNA

    Cre dependent GFP on and RFP off

     

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    Cre efficiently turns off RFP/turns on GFP expression

    HE293 cells were transfected with pAV-CreOn-shRNA alone (Top two panels), or with pAV-CreOn-shRNA and pAV-CAG-Cre.

    Following a 48hr incubation, expression of GFP or RFP was observed by fluorescent microscopy.

    Without the presence of Cre, there were strong RFP expressing cells but no GFP expressing cells. With Cre, there was strong GFP expressing cells but few RFP expressing cells.

     

    Conventional and inducible shRNA expression systems

    Three common methods in shRNA expression:
    1. Conventional shRNA expression with U6 or H1 promoter
    2. MicroRNA scaffolding shRNA expression with polII promoters, such as CMV
    3. Inducible microRNA scaffolding shRNA expression with Tet On promoter



    Search shRNA expression vectors - ready to ship, as lypholyzed 5µg DNA.

    Cre Dependent shRNA expression system

    1. Promoter is flanked by two Loxp and Loxp2722.
    2. Without Cre, promoter drives upstream RFP expression and GFP-microRNA scaffolding shRNA is off.
    3. With Cre, promoter will be flipped, instead of driving RFP, now it drives GFP and microRNA scaffolding shRNA expression.

    Cat# IS100001, $595 three shRNA and one scrambled control.

    pAV-CreOn-shRNA

    ● Cre dependent microRNA scaffolding shRNA expression.
    ● Strong CMV promoter is used in driving shRNA expression and can be replaced by any tissue specific promoters.
    ● RFP can be used to monitor transfection or viral transfection. RFP can be removed from the vector if required.
    ● Loxp and Lox2272 are used in flipping the promoter.
    ● GFP and microRNA scaffolding fusion mRNA can be used as indication of shRNA expression.
    ● Compatible with our AAV, Lentivrial and adenoviral vectors and viral packaging services is available.
    Download pAV-CreOn-shRNA Sequence

     

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