GenJet™ In Vitro DNA Transfection Reagent for HuH-7 Cells

Description
GenJet™ DNA In Vitro Transfection Reagent for HuH-7 Cells is pre-optimized for efficient DNA transfection of human HuH-7 cells.

HuH-7 is a human hepatocellular carcinoma cell line widely used in liver biology, virology, and gene expression studies. Although HuH-7 cells are generally more challenging to transfect than HEK293-derived cells, GenJet™ reagent incorporates proprietary peptide-conjugation technology that enables highly efficient plasmid DNA delivery, achieving transfection efficiencies of up to 70% GFP-positive cells under optimized conditions.

The transfection conditions described below have been optimized to achieve maximal transfection efficiency in HuH-7 cells. One milliliter of GenJet™ reagent is sufficient for approximately 300–600 transfections in 24-well plates or 150–300 transfections in 6-well plates.

Storage Conditions
Store at 4°C. When maintained under recommended storage conditions, the reagent remains stable for at least 24 months.

Transfection Performance of GenJet™ Reagent in HuH-7 Cells

GenJet™ reagent is specifically optimized for HuH-7 cells and demonstrates superior transfection performance compared with Lipofectamine™ 2000 (L2K) and FuGENE® HD. HuH-7 cells were cultured according to recommended conditions and transfected with the GFP expression plasmid pEGFP-N3 using GenJet™ reagent. Transfection efficiency was evaluated 48 hours post-transfection.

Right Panel:
Comparison of transfection efficiency among GenJet™, Lipofectamine™ 2000 (L2K), and FuGENE® HD in HuH-7 cells. Cells were transfected with pEGFP-N3 according to each manufacturer’s recommended protocol. The percentage of GFP-positive cells and relative fluorescence intensity were quantified by flow cytometry (FACS) 48 hours post-transfection.

Left Panel:
The presence of serum and antibiotics enhances GenJet™-mediated transfection efficiency in HuH-7 cells. HuH-7 cells cultured on collagen-coated dishes were transfected under three different conditions: (1) serum- and antibiotic-free medium; (2) medium containing 10% serum and antibiotics, with medium replacement 5 hours post-transfection; and (3) medium containing 10% serum and antibiotics without medium replacement. Transfection efficiency was assessed 48 hours post-transfection.

A representative fluorescence microscopy image demonstrating the transfection efficiency of the pre-optimized GenJet™ Reagent in HuH-7 cells. HuH-7 cells were cultured under recommended growth conditions on collagen type I–coated culture dishes and transfected with the GFP reporter plasmid pEGFP-N3 using GenJet™ Reagent. Transgene expression was assessed 24 hours post-transfection by fluorescence imaging with a Nikon Eclipse 2000 microscope. GFP fluorescence served as an indicator of plasmid DNA delivery and gene expression efficiency.

Transfection Protocol