Constructing an inducible adenovirus system for long non-coding RNA knockdown

Date
2023
DOI
Authors
Liu, Kelly
Version
OA Version
Citation
Abstract
Sickle cell disease (SCD) is an inherited monogenic hemoglobinopathy that affects millions of people worldwide and has been recognized as a global public health problem by the World Health Organization. When sickle red blood cells are damaged and misshaped they tend to block blood flow which can lead to acute vaso-occlusive episodes, hemolytic anemia, stroke, eye retinopathy, and infections. An increase in fetal hemoglobin (HbF) expression can diminish the severity of SCD because HbF can inhibit the polymerization of deoxy-sickle hemoglobin (HbS). In a previous study conducted by the lab, researchers constructed a lentivirus vector to knockdown HBS1L-MYB-intergenic region long non-coding RNA (HMI-lncRNA), which would activate HBG (HBG1 and HBG2) gene expression ultimately increasing HbF synthesis.1 To better understand the role of HMI-lncRNA in regulating HBG, we wanted to have an inducible knockdown system so that we can control the activation and silencing of HMI-lncRNA expression at different time points or development statuses.2 Due to lentivirus vectors having a larger packaging capacity, it gives them a disadvantage in efficient gene delivery compared to adenovirus vectors.3 Therefore, we designed an improved vector to knockdown HMI-lncRNA. This adenovirus vector has a tetracycline-controlled Tet-on/off system incorporated into the short hairpin RNA interference construction. It also carries a green fluorescent protein for selecting and tracking the cells with HMI-lncRNA knockdown. This new inducible adenovirus knockdown system would be a useful tool for our HMI-lncRNA study as well as other lncRNA function studies in other cell lineages.
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