Reporter for Nucleases

Surrogate reporter system


Targeting efficiency of Engineered nucleases ranges 1%~30% for the most cases. Although this efficiency is quite remarkable compared to conventional gene targeting techniques by homologous recombination, steps to isolate cells bearing desired mutations still require extensive screening.

Our knockout cell enrichment system has been developed to overcome such limitations. The use of surrogate reporters allows selective enrichment of cells that have acquired EN mediated mutations at target sites.

The surrogate reporters are composed of genes encoding two fluorescent proteins (red and green) linked by the EN targeting sequence. In the absence of EN expression, they are designed to express the red but not the green fluorescent protein as it is placed out of frame. Upon expression of ENs, which can induce double strand break at the target site in front of GFP, leading to frameshift mutations and expression of GFP. The expression of green GFP is strictly dependent on the presence of nuclease activity from ENs that specifically recognize the target sequence in the reporter construct.


RGN CRISPR/Cas9

Our surrogate reporter system is a wonderful tool to visualize and enrich the cells with gene editing event. Using surrogate reporter system, the frequency of finding the knock out cells can be improved by 3~20 fold.

RGN CRISPR/Cas9

Surrogate Reporter Systems

  • GFP system: mRFP (constitutive) + EN targeting site + GFP1 (out of frame) + GFP2 (out of frame)
  • MACS system: mRFP (constitutive) + EN targeting site + GFP (out of frame) + transmembrane protein (out of frame)
  • HygR system: mRFP (constitutive) + EN targeting site + GFP (out of frame) + Hygromycin-R (out of frame)
  • Time line: 2 weeks


  • RGN CRISPR/Cas9


    Cas9 co-transfection/selection vectors

  • CV02: Cas9, Hygro R, and GFP (CMV promoter)
  • CV03: Cas9, RFP, and Puro R (CMV promoter)
  • CV12: Cas9, Hygro R, and GFP (EF1a promoter)
  • CV13: Cas9, RFP, and Puro R (EF1a promoter)


  • Cat No
    Item
    Description
    Size
    Price
    Order
    RV01
    pRGS

    Surrogate reporter vector with GFP for FACS

    5 ug
    $350.00
    RV02
    pMRS

    Surrogate reporter vector with GFP and H2KK for MACS

    5 ug
    $350.00
    RV03
    pHRS

    Surrogate reporter vector with GFP and Hygromycin-R

    5 ug
    $350.00
    CV01
    Cas9_CMV

    Cas9 co-transfection vector (CMV promoter)

    5 ug
    $250.00
    CV02
    Cas9_HygR_GFP (CMVp)

    Cas9, Hygromycin R, and GFP expression vector (CMV promoter)

    5 ug
    $350.00
    CV03
    Cas9_RFP_PuroR (CMVp)

    Cas9, RFP, puromycin R expression vector (CMV promoter)

    5 ug
    $350.00
    CV11
    Cas9_EF1a

    Cas9 co-expression vector (EF1a promoter)

    5 ug
    $350.00
    CV12
    Cas9_HygR_GFP (EF1a)

    Cas9, Hygromycin R, and GFP expression vector (EF1a promoter)

    5 ug
    $350.00
    CV13
    Cas9_RFP_PuroR (EF1a)

    Cas9, RFP, and Puromycin R expression vector (EF1a promoter)

    5 ug
    $350.00
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