TcBuster™ Transposon CD19CAR-DHFR-eGFP

Name: TcBuster™ Transposon CD19CAR-DHFR-eGFP
Brand: R&D Systems
SKU: TCBP001-100
Price: 1300 USD
Availability: InStock

R&D Systems | Catalog # TCBP001-100

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Product Documents

DNA transposon component of the TcBuster non-viral gene delivery system.

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Product Summary
Scientific Data
Preparation & Storage
Product Documents
Specific Notices

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

Product 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 EF1 alpha 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.

Scientific Data Images for TcBuster™ Transposon CD19CAR-DHFR-eGFP

TcBuster Transposon CD19CAR-DHFR-eGFP Plasmid Map.

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).

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 CD19CAR-DHFR-eGFP

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Certificate of Analysis

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Product Specific Notices for TcBuster™ Transposon CD19CAR-DHFR-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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