HSDL1 Knockout HEK293T Polyclonal Cells are a mixed population of HEK293T cells bearing CRISPR/Cas9-mediated disruption of the HSDL1 gene. Unlike monoclonal isolates, this polyclonal format captures a spectrum of knockout alleles, enabling population-level functional analysis. The heterogeneous gene disruption mimics physiological variation and is ideal for initial characterization of HSDL1 loss-of-function phenotypes in an epithelial cell background. This product provides a cost-effective and reproducible foundation for mitochondrial and lipid metabolism research.
HEK293T cells are an immortalized human embryonic kidney epithelial line stably expressing SV40 large T antigen, which enhances episomal replication of transfected plasmids and supports high-level recombinant protein expression. They retain core mitochondrial and peroxisomal pathways, making them a relevant host for studying lipid and steroid metabolism. The epithelial origin and robust transfection efficiency enable mechanistic dissection of HSDL1 function via overexpression, knockdown, and rescue experiments in a controlled setting.
HSDL1 encodes a putative mitochondrial short-chain dehydrogenase/reductase that catalyzes NAD+-dependent oxidation/reduction of hydroxysteroids and fatty acid intermediates. Its expression is transcriptionally activated by nuclear receptors PPAR?? and LXR??, and by SREBP1, linking HSDL1 to lipid and cholesterol homeostasis. HSDL1 interacts with mitochondrial SDR family members and STAR protein, and promotes expression of fatty acid oxidation genes such as ACADM and CPT1A. It also influences steroidogenic enzymes including HSD3B1, CYP17A1, and AKR1C3. Thus, HSDL1 integrates mitochondrial fatty acid ??-oxidation and steroid biosynthesis, with knockout predicted to impair cellular energy and lipid metabolism.
Knockout of HSDL1 in HEK293T cells creates a versatile model for studying mitochondrial lipid and steroid metabolic reprogramming in a human epithelial context. Though HEK293T cells are not steroidogenic, they retain metabolic sensors and can be engineered to express relevant regulators. The polyclonal knockout pool allows analysis of mitochondrial respiration, lipid storage, and crosstalk between AMPK and PPAR??. High transfection efficiency enables rescue with wild-type or mutant HSDL1 and real-time biosensor monitoring of lipid flux, while the polyclonal nature reflects population heterogeneity observed in RNAi screens.
Key applications include Seahorse respirometry for mitochondrial fatty acid oxidation, LC-MS for steroid profiling, and lipid droplet staining for storage assessment. Transcriptomic analysis by RNA-seq and validation by RT-qPCR/Western blotting define downstream networks, while flow cytometry for mitochondrial mass gauges adaptive biogenesis. These cells are suited for drug screens targeting PPAR??, LXR??, or SREBP1 to bypass HSDL1 deficiency. For further information, contact Ascent Research.
