This product consists of a polyclonal HeLa cell population engineered using CRISPR/Cas9 to disrupt the DROSHA gene, generating a heterogeneous pool of knockout cells. The polyclonal format provides a robust loss-of-function model without the clonal selection artifacts that can arise in single-cell?Cderived lines. Each vial contains a diverse cell population with targeted disruption of DROSHA, allowing researchers to study the consequences of impaired primary microRNA processing in a physiologically relevant cellular context.
The parental HeLa cell line is an immortalized human cervical adenocarcinoma epithelial cell line originally derived from Henrietta Lacks in 1951. HeLa cells are among the most widely used model systems in biomedical research, offering robust growth characteristics, ease of transfection, and a well-characterized cancer cell background. Their epithelial origin and transformed nature make them especially relevant for studies of cancer biology, gene regulation, and host?Cpathogen interactions.
DROSHA encodes the catalytic subunit of the Microprocessor complex, which also includes the double?stranded RNA-binding protein DGCR8. In the nucleus, this complex recognizes and cleaves primary microRNA (pri?miRNA) transcripts to release ~70?nt stem?loop precursor microRNAs (pre?miRNAs), initiating the canonical miRNA biogenesis pathway. This activity is regulated by upstream signaling factors including the tumor suppressor p53, the oncoprotein MYC, SMAD2/3, and ERK signaling cascades. Processed pre?miRNAs are subsequently exported to the cytoplasm by Exportin-5 and further matured by Dicer, TRBP, and AGO2 to form the RNA-induced silencing complex (RISC), which mediates post?transcriptional repression of target mRNAs. DROSHA activity thus influences the expression of numerous mature microRNAs, including key cancer?associated species such as let?7, miR?21, and miR?155. Additionally, DROSHA interacts with several RNA-binding proteins (e.g., DDX5, DDX17, FUS, HNRNPA1) and SR proteins that modulate Microprocessor activity and substrate selectivity.
In the HeLa cervical adenocarcinoma context, DROSHA knockout disrupts the global microRNA landscape, providing a powerful model to dissect microRNA?dependent mechanisms of gene silencing and their contributions to malignant phenotypes. Because HeLa cells express a wide array of oncogenic and tumor?suppressive microRNAs, loss of DROSHA enables systematic investigation of how pri?miRNA processing defects alter cancer cell proliferation, apoptosis, migration, and drug sensitivity. This model is particularly valuable for studying the interplay between miRNA biogenesis and pathways frequently dysregulated in cervical cancer and other malignancies.
Researchers can employ these polyclonal knockout cells in a variety of functional and mechanistic assays. miRNA expression profiling by small RNA?sequencing or RT?qPCR can quantify the global loss of mature miRNAs, while Microprocessor activity assays and pri?miRNA processing assays directly assess the block in miRNA maturation. Co?immunoprecipitation of the DROSHA?DGCR8 complex, RNA immunoprecipitation (RIP), and western blotting for downstream target proteins can elucidate changes in miRNA?mediated regulation. Luciferase reporters carrying miRNA target sites can functionally validate altered silencing activity. These cells are thus suited for miRNA biogenesis studies, cancer biology, gene silencing research, functional genomics screens, and drug target discovery efforts. For any inquiries about this product, please contact Ascent Research.
