TcBuster™ Transposon Insert On eGFP 

R&D Systems | Catalog # TCBP002-100

DNA transposon component of the TcBuster non-viral gene delivery system.
R&D Systems
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Key Product Details

Features

TcBuster Transposon Insert On eGFP is a DNA transposon (plasmid) containing an eGFP sequence flanked by TcBuster-compatible ITR sequences that is only expressed after genomic integration. As a component of the TcBuster non-viral gene delivery system, this product requires TcBuster-M Transposase mRNA to function. This two component system can be introduced, by electroporation, into a wide variety of cell types to enable stable gene transfer.

Key Benefits

  • Used as an eGFP-expressing control for the TcBuster system in both cell therapy and bioproduction applications
  • Based on a genetic switch, expression of eGFP only occurs after transposition and is a direct readout of cargo (eGFP) integration

Product Summary for TcBuster™ Transposon Insert On eGFP 

What is the TcBuster Non-Viral Gene Delivery System?
Gene delivery is a crucial step for a wide variety of genome engineering workflows, which have traditionally utilized viral vectors to achieve expression of exogenous DNA sequences. However, the generation of a virus is time consuming, inconsistent, and has limited cargo capacity. The TcBuster non-viral gene delivery system is a solution for stable integration of DNA cargo and addresses many of the limitations of viral vectors. It consists of the TcBuster-M Transposase mRNA and a compatible DNA TcBuster transposon, such as TcBuster Transposon Insert On eGFP. This product requires TcBuster Transposase mRNA to be functional.
Why use TcBuster Transposon Insert On eGFP?
This transposon encodes DNA cargo consisting of an eGFP sequence regulated by the EF1 alpha promoter. It also incorporates a genetic switch, called integration-coupled On, that effectively eliminates expression of the untransposed plasmid (1). Gene expression can only occur after transposition and is a direct readout of cargo integration. When used in combination with TcBuster-M transposase mRNA, the DNA cargo will be inserted randomly into the host cell genome. This transposon serves as an ideal DNA cargo for initial evaluations of the TcBuster system and/or a control for cell therapy and bioproduction applications using the TcBuster system.

(1) Kumamoto, T. et al..2020. Neuroresource

Scientific Data Images for TcBuster™ Transposon Insert On eGFP 

Integration-coupled genetic switch composed of an EF1a promoter regulating the expression of eGFP

TcBuster Transposon Insert On eGFP Plasmid Map.

HEK293E after electroporation with TcBuster Transposon Insert On eGFP. When electroporated with TcBuster-M, eGFP expression is measurable by flow cytometry, while the absence shows no expression from the unintegrated transposon.

HEK293E cells edited with the TcBuster system express eGFP.

HEK293E cells were gene edited with TcBuster-M transposase (Catalog # TCB-001.1-100) and Insert-On-eGFP transposon (Catalog # TCBP002-100). On day 2 after gene editing, cells were analyzed by flow cytometry for expression of eGFP. Representative flow cytometry plots of HEK293E cells edited with or without TcBuster-M transposase.

CHO-K1 after electroporation with TcBuster Transposon Insert On eGFP and with or without TcBuster Transposase mRNA, showing no expression from the unintegrated transposon.

CHO-K1 cells edited with the TcBuster system express eGFP.

CHO-K1 cells were gene edited with TcBuster-M transposase (Catalog # TCB-001.1-100) and Insert-On-eGFP transposon (Catalog # TCBP002-100). On day 2 after gene editing, cells were analyzed by flow cytometry for expression of eGFP. Representative flow cytometry plots of CHO-K1 cells edited with or without TcBuster-M transposase.

Formulation, Preparation, and Storage

Shipping

This product ships at ≤ -20 °C. Upon receipt, store it immediately at the recommended temperature.

Storage

Store at ≤ -20 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

Product Documents for TcBuster™ Transposon Insert On eGFP 

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for TcBuster™ Transposon Insert On eGFP 

For laboratory research use only. Not for use in diagnostic procedures.

  • For certain applications, results may vary due to differences in donor-derived cell populations.

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