The DHRS1 Knockout HEK293T Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DHRS1 gene in HEK293T human embryonic kidney cells. This loss-of-function model disrupts the gene encoding dehydrogenase/reductase SDR family member 1, a key enzyme in retinoid metabolism. The polyclonal format provides a genetically diverse pool of edited cells, enabling bulk-population studies without clonal selection bias, suitable for a wide range of functional assays investigating DHRS1-dependent processes.
The parental HEK293T cell line, transformed with adenovirus 5 DNA and stably expressing SV40 large T antigen, supports episomal replication of SV40 origin plasmids and exhibits exceptionally high transfection efficiency. Widely used for recombinant protein production and viral packaging, HEK293T cells offer a robust and well-characterized platform for generating gene knockouts, facilitating the dissection of specific gene functions in a human cellular context.
DHRS1 functions as an NADPH-dependent short-chain dehydrogenase/reductase that catalyzes the reduction of all-trans-retinal to all-trans-retinol, a critical step in retinoid cycling that modulates intracellular retinoic acid pools. Its activity is regulated by retinoic acid and oxidative stress via NRF2, and its product feeds into RAR/RXR nuclear receptor signaling programs controlling cell differentiation and proliferation. Within the retinoid metabolic network, DHRS1 interacts with NADPH cofactor and cellular retinol-binding proteins (CRBPs) and operates alongside RDH5, RDH10, and ALDH1A1, linking enzymatic retinoid processing to downstream transcriptional regulation of differentiation genes.
In the HEK293T background, DHRS1 knockout creates a model to examine retinoid metabolism independent of endogenous differentiation constraints, as these kidney-derived cells do not rely on retinoid signaling for survival. The polyclonal population captures heterogeneous editing outcomes, reflecting physiologically relevant variability in gene function. Combined with the high transfectability of HEK293T cells, this model is ideal for complementation studies, overexpression of pathway components such as RAR?? or CRABP1, and downstream reporter assays to map DHRS1-dependent signaling. This system enables dissection of how DHRS1 loss alters metabolite levels and gene expression without confounding differentiation programs.
Key applications include retinoid metabolite quantification by LC-MS, proliferation assessment via MTT assays, and differentiation marker analysis by RT-qPCR or western blotting. The knockout cells are also suitable for drug screening targeting retinoid pathways in cancer or metabolic diseases, enabling high-throughput chemical screens and target validation through rescue experiments. These cells facilitate investigations into crosstalk between retinoid metabolism, oxidative stress, and oncogenic signaling. Researchers can further utilize the polyclonal format to study the functional impact of DHRS1 disruption on cellular retinoic acid responses and phenotypic plasticity. For custom inquiries, please contact Ascent Research.
