Critical factors for lentivirus-mediated PRDX4 gene transfer in the HepG2 cell line.

ABSTRACT: Optimization of critical factors affects transduction efficiency and is able to reduce reagent consumption. The present study aimed to determine the optimum transduction conditions of small hairpin (sh)RNA against peroxiredoxin 4 (PRDX4) in the HepG2 cell line. Cell viability assays were conducted based on serum condition, incubation time, polybrene concentration and antibiotic dose selection. Non-targeting control shRNA was transduced into HepG2 cells in a 5-fold serial dilution, and colonies positive for green fluorescent protein were counted using ImageJ software. Reverse transcription-quantitative polymerase chain reaction and western blot analysis were performed to validate PRDX4 expression. The optimum cell density for transduction was 5.0×10³ cells/well in 96-well plates

SUBMITTER: Aznan AN 

PROVIDER: S-EPMC6006497 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature