The DROSHA Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T human embryonic kidney epithelial cells, harboring a targeted disruption of the DROSHA gene. This loss-of-function model enables study of the microprocessor complex and miRNA biogenesis in a well-characterized host. The polyclonal pool comprises a heterogeneous collection of edited alleles, ensuring robust knockout without clonal isolation, suitable for population-level analyses.
HEK293T cells are widely used for protein expression and viral production due to their adenovirus 5-transformed origin and constitutive SV40 large T antigen expression, which boosts plasmid replication. Their epithelial nature and high transfectability make them an ideal platform for investigating RNA processing and gene regulation mechanisms.
DROSHA is a ribonuclease III that partners with DGCR8 to cleave pri-miRNAs into pre-miRNAs in the nucleus, initiating miRNA biogenesis. The resulting pre-miRNAs are exported by Exportin-5 and processed by Dicer into mature miRNAs that assemble into RISC for target mRNA silencing. DROSHA activity is regulated by p53 and TGF-??/SMAD signaling, and its interaction with DGCR8 is stabilized by DEAD-box helicases (DDX5, DDX17), hnRNPs, and FUS/TLS. Among its downstream targets are miRNAs let-7, miR-21, and miR-155, which govern cell fate decisions.
Knocking out DROSHA in HEK293T cells eliminates canonical miRNA production, leading to pri-miRNA accumulation and loss of miRNA-directed gene regulation. This system facilitates transfection-based rescue experiments and mechanistic dissection of microprocessor function. The SV40 large T antigen background does not interfere with miRNA processing, providing a clean context for studying DROSHA-dependent phenotypes.
Applications include small RNA sequencing to profile miRNA changes, pri-miRNA processing assays, co-immunoprecipitation of microprocessor components, and luciferase reporter assays to measure miRNA activity. The cells are also valuable for screening miRNA biogenesis modulators and exploring DROSHA??s role in cancer and developmental biology. For inquiries, contact Ascent Research.
