HyCyte™ Gene Interference Stabilization Kit

The second-generation HyCyte™ Gene Interference Stabilization Kit (shRNAx2.0) is a gene interference stabilization kit developed based on site-specific SSR recombinase technology.

Product Features:
Utilizing SSR (Site-specific Recombinase) recombinase to achieve highly efficient and stable transfection of interference sequence vectors, significantly improving interference efficiency. The transfection procedure is the same as siRNA transfection, constructing a gene interference stable transfection cell line similar to a virus, making it a next-generation gene interference technology product.

The interference kit includes all genes in the human and mouse genomes. The shRNAx2.0 kit contains two selected shRNA interference sequence vectors, one GFP/control shRNA interference sequence vector, and one tube of site-specific SSR recombinase. The SSR recombinase efficiently catalyzes the integration of the shRNA vector into the cell genome, achieving stable transfection of the shRNA fragment. Simultaneously, due to the extremely high integration efficiency of SSR recombinase, the shRNA fragment contains approximately 10-100 copies in the target cell genome. This high copy number significantly enhances the expression of the shRNA fragment, thereby improving the gene interference effect. The shRNAx2.0 vector contains a puro resistance gene and a GFP fluorescent gene, allowing for easy assessment of the transfection efficiency of the interference vector and the expression efficiency of the target vector via fluorescence. High-expression cell lines can be obtained through Puromycin screening, facilitating the establishment of stable gene interference cell lines. The shRNAx2.0 interference stabilization kit is as simple to use as the siRNA transfection process, exhibiting excellent interference effects on various cell types and is also suitable for gene function studies in mesenchymal stem cells (MSCs).

Product Advantages
· Simpler than viral methods – excellent interference effects on both dividing and non-dividing cells.
· High cell stabilization efficiency – long-term stable interference cell lines can be obtained in just 1-2 weeks.

Application Procedure
· Liposome Transfection:
This kit can use Lipofectamine® 2000 Reagent. Refer to the specific transfection reagent for each cell type. For different types of transfection reagents, please follow the manufacturer's instructions for transfection.
Note: Liposome transfection is not recommended for suspension cells.
· Electroporation:
This kit is compatible with the Neon transfection system. Refer to the equipment specifications for each cell type. For different types of electroporation equipment, please optimize according to the manufacturer's instructions.
Note: Electroporation is recommended for suspension cells.

The HyCyte™ Gene Interference Stabilization Kit (shRNAx2.0) includes a GFP control shRNA vector. Experiments can be conducted based on the actual method and cell conditions. For optimal results, the GFP positivity rate of the target cells should be at least 50% after 48 hours.

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Background

The HyCyte™ Gene Interference Stabilization Kit (shRNAx2.0) offers interference efficiency comparable to traditional viral methods. Its unique SSR recombinase enables the construction of high-copy "stable" cell lines, and efficient, stable interference-mediated cell lines can be screened using drugs. The shRNAx2.0 kit is the only experimental protocol besides viral methods that can easily obtain stable interference-mediated cell lines.
Using the HyCyte™ Gene Interference Stabilization Kit (shRNAx2.0), the construction of stable interference-mediated cell lines takes only 1-2 weeks, providing long-term stable interference, significantly accelerating experimental progress.