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TcBuster™ Non-Viral Gene Engineering

The TcBuster Transposon Gene Editing Mechanism

TcBuster is a large capacity transposon system that enables transfer and stable integration of genes of interest (GOI). TcBuster can replace the use of retroviral and lentiviral vectors, leading to significant time savings during the development of cell therapeutics.

Why Use Non-Viral Vectors For Gene Engineering?

TcBuster is a non-viral transposon system that enables stable gene transfer into mammalian cells. It supports the rapid generation of transgenic cells and is a valuable tool for both proof-of-concept and manufacturing scale process development. Gene editing with TcBuster has widespread applications including:

  • Cancer immunotherapies – introduction of chimeric antigen receptors (CARs) or TCRs; inactivation of negative regulatory molecules; masking therapeutic cells from host immunity
  • Gene therapy – delivery of genes into host cells to correct genetic defects
  • Bioprocessing - significantly improve bioprocessing yields
  • Disease models – development of animal models carrying disease-associated molecular variants
  • Drug discovery – modulating expression of proteins that serve as drug targets
  • Diagnostic tools – upregulation of proteins for detection of infectious and non-infectious diseases
Gene Engineering With TcBuster

Comparing TcBuster and Virus-Based Systems

Sidestep the hurdles of gene engineering with lentivirus systems by using a non-viral method. The use of TcBuster avoids common viral method challenges including low transduction efficiency, inconsistent reagent availability, and an insecure supply chain for GMP-grade virus.

Virus-based gene delivery (e.g. with lentivirus or AAV) involves packaging of the GOI into the viral vector followed by transduction of target cells. TcBuster transposons can be quickly generated and electroporated into target cells. As shown in the graphic below, TcBuster protocols eliminate the lengthy step for viral vector packaging.

Enhance your cell engineering process with TcBuster and:

  • Shorten your cell engineering timeline and reduce the risk of delays.
  • Integrate larger cargo sizes than with standard viral based methods. AAV can typically deliver 4-5 kb of DNA, while TcBuster can deliver >10 kb (enough for 2 TCRs plus an accessory protein).
  • Maintain high efficiency gene transfer and stable expression. This enables rapid generation of transgenic mammalian cells with limited secondary effects and no selection steps.
  • Reduce the costs of genetic modification.
  • Exploit the flexibility of TcBuster to carry out additional gene editing techniques in one operation.
  • Scale from research to clinical and commercial effortlessly with TcBuster gene editing and our partners at ScaleReady
timeline of gene engineering with TcBuster compared to virus-based systems

Hear From Bio-Techne Scientists

Take your cell therapy beyond the payload capacity limit with a non-viral transposon system.

Watch this video to hear from our experts about gene engineering with TcBuster.

Realize the potential of gene engineering by using TcBuster to deliver large genes of interest. Gene editing is a powerful tool for fine tuning cell phenotype in immune cell and regenerative medicine therapies, and it can augment therapeutic effectiveness by increasing cell potency and persistence in vivo.

Partner with our experts to engineer T cells, NK cells, feeder cells, and induced pluripotent stem cells (iPSC) for your cell therapy program.

Transfection Efficiency with Non-Viral Gene Delivery

(Left) Transfection efficiency of T cells using TcBuster for CD19 CAR, (Right) Killing efficiency of CAR-T cells generated with TcBuster

Similar CAR expression was achieved in primary human T cells edited with a CD19 CAR using TcBuster or lentivirus as shown by detection with the Fluorokine fluorescent CD19 antigen (left 2 panels). In the right panel, CD19 CAR-T cells engineered using TcBuster (dark blue) or lentivirus (light blue) show similar cytotoxicity against CD19+ Nalm6/luc cells. The non-electroporated controls (gray) do not kill target cells. 

Delivery of large CAR constructs with TcBuster

Transposition Capacity With TcBuster

TcBuster can efficiently deliver both small single CAR constructs and large-sized, multi-cistronic constructs. Activated T cells were transposed with increasing plasmid sizes, expanded for 4 days, and analyzed for CAR expression by flow cytometry. >30% transposition efficiency was achieved with constructs ranging up to 6 kb of insert. 

Steps In Partnering With Us

Realize the potential of gene engineering efficiently by partnering with our expert team. Whether you are new to the technique or represent a gene engineering core group with limited bandwidth, we will provide the same level of responsive partnership and customer service. We will connect scientist-to-scientist and walk through the entire project with you.

Cell therapy gene engineering with TcBuster gene transfer system

Project scope and experimental design

  • Timelines, milestones, deliverables, and project team
  • Custom requirements for deliverables
  • Timely quote delivery

What to expect throughout the project

  • Accessibility of project manager or technical lead to facilitate discussions
  • Delivery of technical progress reports as defined in the statement of work (SOW)
  • Shipment of cell samples and other materials for your testing

What defines project completion?

  • Shipment of engineered cells in agreed-upon formulation, packaging, and labeling
  • Delivery of characterization reports, documentation, and other deliverables as defined in the SOW
  • Quarantine of extra materials
  • Project offboarding and archiving

Material Transfer Agreements (MTAs)

  • Perform cell engineering with TcBuster in your own lab
  • Research licenses to support long term discovery programs
  • Commercial licenses for both therapeutics and ancillary materials
  • Available to qualified laboratories

Bio-Techne believes in the power of partnerships. We have joined with Fresenius Kabi and Wilson Wolf to bring you ScaleReady™, lowering the barriers to entry into the immune cell therapy space for all sizes and stages of manufacturing programs. ScaleReady brings cell and gene therapies to life by offering a significant reduction of complexity and cost while consistently providing superior repeatability and cell quality.

Partner with ScaleReady for TcBuster applications in immune cell therapy. For other applications of TcBuster, contact a gene engineering expert at Bio-Techne.

Luminary Therapeutics Corporate Logo

Luminary Therapeutics has signed a licence agreement for the use of TcBuster for the manufacturing of gene-modified cell therapies. Luminary Therapeutics will use TcBuster in their LMY-920 program, a BAFF-CAR-T product.

Cell & Gene Therapy article on TcBuster

This article in Cell & Gene Therapy Insights discusses the transposon-based TcBuster platform and its competitive advantages in cell and gene therapy. Directed evolution of hyperactive TcBuster, TcBuster in cell therapies, advantages and applications of TcBuster, Q&A with the authors.

T Cell-Based Therapies eBook

This eBook provides an overview of several biological and manufacturing challenges facing the development and production of T cell therapies. It follows with examples of how Bio-Techne products and services can address these challenges at each stage of the process.

Frequently Asked Questions

Tell us about your project, and one of our experts will contact you to discuss how we can help.