DNA transposon component of the TcBuster non-viral gene delivery system.
Key Product Details
Features:
TcBuster Transposon CD19CAR-DHFR-eGFP is a DNA transposon (plasmid) containing CD19CAR, DHFR mutein, and eGFP sequences flanked by TcBuster-compatible ITR sequences. 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:
Contains a large DNA cargo insert (5.1 kb) for use in the TcBuster system
Used as a control for cell therapy applications, as this transposon encodes for a CD19 CAR, DHFR mutein, and eGFP sequences
Built with a minimal backbone (Nanoplasmid™) for efficient electroporation
Summary for TcBuster™ Transposon CD19CAR-DHFR-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 CD19CAR-DHFR-eGFP. This product requires TcBuster Transposase mRNA to be functional.
Why use TcBuster Transposon CD19CAR-DHFR-eGFP?
This transposon encodes a multicistronic DNA cargo consisting of a CD19CAR, DHFR mutein, and eGFP sequence regulated by the EF1α promoter. 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 applications using the TcBuster system.
Limitations
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.
Scientific Data Examples for TcBuster™ Transposon CD19CAR-DHFR-eGFP
Primary human T cells edited with the TcBuster system express CD19CAR and eGFP.
Primary human T cells were gene edited with TcBuster-M transposase (Catalog # TCB-001.1-100) and CD19CAR-DHFR-eGFP transposon (Catalog # TCBP001-100). On day 7 after gene editing, T cells were stained with Recombinant Human CD19 Protein, Atto 647N Conjugate (Catalog # ATM9269) and analyzed by flow cytometry for CD19CAR and eGFP expression. Representative flow cytometry plots of primary T cells edited with or without TcBuster-M transposase.
Mechanistic Diagram of the TcBuster Non-Viral Gene Editing System.
The TcBuster system components are introduced into cells via electroporation (1). The TcBuster-M transposase mRNA is then translated into the transposase enzyme (2), which binds to the inverted terminal repeats (ITRs) on the DNA transposon (3). The transposase enzyme excises the genes of interest (GOI) from the transposon (4) and inserts them into the host genome (5). The GOI mRNA is transcribed from the host genome (6), and the protein is stably expressed in the edited cells (7).
Preparation & Storage
Shipping Conditions
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.
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