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Leap-In Transposases®

    Expression from pD2500 Leap-In Stable Vectors can be further increased by co-transfecting with one of our new transposases. Many pD2500 vectors already perform extremely well for expression of single proteins, even without transposase. Transposases are potent tools for eliminating yield reductions that can otherwise occur in stable cell lines containing IRES-based bicistronic constructs. Use in combination with our glutamine synthase or dihydrofolate reductase markers to boost antibody production even faster.

    Leap-In Transposase Increases Protein Expression


    Transposase Protein Expression
    Fluorescent protein expression compared in CHO cell populations transfected with and without transposase. CHO cells were transfected with pD2500 vectors carrying a gene for DasherGFP, with (green bars) or without (blue bars) a plasmid carrying a transposase gene. Cells were harvested for DasherGFP fluorescence measurements after 14 days (2 passages post-selection) from stably transfected CHO cells.
    *piggyBac® is a registered trademark of Transposagen, Inc.

    Leap-In Transposase Increases Bicistronic Protein Expression


    Transposase bicistronic protein expression
    Bicistronic fluorescent protein expression compared in CHO cell populations transfected with and without transposase. CHO cells were transfected with pD2539 carrying DasherGFP-IRES-CayenneRFP, with (green bars) or without (blue bars) a plasmid carrying a transposase gene. Cells were harvested for fluorescence measurements after 14 days (2 passages postselection) from stably transfected CHO cells.

    FACS Analysis: Cell Populations Shift From Poor to High Expression


    Transposase FACS expression results
    DasherGFP expression compared in CHO cell populations transfected with and without transposase. CHO cells were transfected with pD2500 vectors carrying a gene for DasherGFP, with or without a plasmid carrying a transposase gene. Cells were harvested for GFP FACS analysis after 14 days (2 passages post-selection) from stably transfected CHO cells.

    Applications

    • Protein expression
    • Genome editing

    Mechanism

    The Leap-In Transposase is an engineered enzyme that catalyzes the integration of a transposon, your gene, into TTAT sites in the target genome. The technology enables a specified sequence to behave as a transposon, a mobile genetic element, which can efficiently transpose between vectors and chromosomes via a “cut and paste” mechanism. During transposition, the Leap-In Transposase recognizes transposon-specific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and moves the contents from the original sites and efficiently integrates them into TTAT chromosomal sites. Similar technologies report integration of up to 20 copies of the gene into unique locations in the genome 72 hours post transfection.

    Transposase Mechanism
    A gene between Inverted Terminal Repeats (ITR) carried by a plasmid vector is transfected into the cell with the transposase, as mRNA, protein, or expressed from a plasmid. The gene is excised from the donor plasmid at the Transposon Recognition Sites (TRSs) and multiple copies are integrated into TRSs found in the chromosomal DNA.

    Advantages

    • Up to 20 stable copies of your gene integrated into your target genome 72 hours post transfection
    • Stable pools created in 2 weeks, Cell-Lines 6 weeks
    • Single Transfection
    • Unlimited payload

    ATUM Commercial Licensing

    Please contact us for licensing terms for research and commercial applications at info@atum.bio.


    Consult an Expression Expert

    To determine the best possible option for your research project contact us at: +1 877 362 8646 or info@atum.bio.