Peptide-based nucleic acid delivery is a state-of-the-art delivery technology that can significantly improve nucleic acid transfection of a wide range of cells, including primary cells and hard-to-transfect cell lines. Based on a high-throughput peptide screening platform, GentleFectTM technology accurately identifies endogenous peptide sequences with phase separation potential, and developed the Gentle T30 nucleic acid transfection reagent for primary T cells. Peptides are self-assembled by liquid-liquid phase separation with nucleic acids to form nano-scale droplets, which can be efficiently internalized with the help of cell membrane pinocytosis, resulting significantly improvement of the nucleic acid delivery efficiency of cells across all species, while cell viability are also maintained.
• The product is suitable for transfection of various nucleic acid molecules, including mRNA, siRNA, circRNA, Crispr, etc.;
• The product is a ready-to use transfection reagent, easily add nucleic acids to the reagent to configure the transfection solution in a single step.
4°C for short storage and -20°C for long storage.
• Gentle T30 Nucleic Acid Transfection Reagents for primary T cells
No, due to their unique immune characteristics, primary T cells are sensitive to exogenous double-stranded DNA by the intracellular cGAS-STING signaling pathway, which not only causes cell cycle arrest, but also leads to cell death through the endogenous apoptotic pathway, and the cell viability and positivity rate will be significantly reduced under conventional transfection conditions. See Reference: Enhancement of the viability of T cells electroporated with DNA via osmotic dampening of the DNA-sensing cGAS–STING pathway. Nature Biomedical Engineering.
Primary T cells are suspension cells, this reagent can achieve efficient and low-toxicity transfection of primary T cells in a 96-well plate format, covering a wide range of cell numbers from 1x104 to 2x105. It is recommended to follow the transfection reagent amounts specified in Instruction manual . Alternatively, the amount of transfection reagent can be adjusted proportionally according to the number of cells, with a minimum requirement of 5x103 cells.
① Optimize Cell Condition: Prior to transfection, ensure cells viability exceed 90%. This can be assessed using standard viability assays such as trypan blue exclusion or propidium iodide staining.
② Adjust Reagent Concentrations: Incrementally increase the amounts of transfection reagent and nucleic acid to identify the optimal ratio that maximizes transfection efficiency without compromising cell viability.
③ Optimize Incubation Time: Experiment with varying incubation times to determine the ideal duration for your specific cell type. Both prolonged and shortened incubation periods can influence transfection efficiency and cell viability, so it is essential to find a balance that yields the best results.
① Do not use medium containing FBS during cell transfection, FBS will significantly compromise the efficiency of cell transfection;
② Gentle T30 Transfection Reagent is not recommended to vortex vigorously, and it is recommended to use a pipette to mix by gentle and repeated pipetting.
Gentle T30 Nucleic Acid Transfection Kit for Primary T Cells v1.3
