The ASCC3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ASCC3 gene in a mixed clonal background. This product provides a pooled knockout model generated by CRISPR/Cas9-mediated gene disruption, suitable for studying loss-of-function phenotypes without the constraints of single-cell clonal selection. The polyclonal format preserves heterogeneous editing outcomes, offering a convenient approach to assess bulk population effects of ASCC3 deficiency.
HEK293T cells are a widely utilized human embryonic kidney epithelial cell line immortalized by the SV40 large T antigen. This transformation confers robust proliferation and high transfection efficiency, making HEK293T a preferred host for genetic manipulation and biochemical assays. The epithelial origin and stable karyotype support investigations of DNA damage response, transcription regulation, and cancer-related pathways.
ASCC3 encodes an ATP-dependent DNA helicase that serves as the catalytic subunit of the ASC-1 complex, which also includes ASCC1, ASCC2, and TRIP4. This complex is recruited to sites of transcription-blocking DNA lesions, where ASCC3 unwinds DNA secondary structures and resolves R-loops to facilitate transcription restart. Its activity is triggered by upstream signals such as stalled RNA polymerase II and ATR kinase activation following DNA damage from UV radiation or alkylating agents. ASCC3 functionally interacts with the transcription-coupled repair factors ERCC6 (CSB) and ERCC8 (CSA), and it mediates the recruitment of the alkylation repair enzyme ALKBH3 to damaged chromatin. Downstream consequences of ASCC3 function include restored mRNA synthesis, removal of DNA alkylation lesions, and efficient RNA polymerase II transcription elongation.
In the HEK293T background, disruption of ASCC3 creates a valuable model for dissecting the interplay between transcription and DNA repair. Given the high transcriptional activity of these cells, ASCC3 loss is expected to impair resolution of R-loops and hinder transcription-coupled repair processes. This knockout cell population enables the study of endogenous ASCC3 functions without the confounding effects of clonal variation, allowing researchers to probe pathway dynamics using standard molecular and cellular assays.
This polyclonal knockout model is well-suited for a range of experimental applications, including mechanistic studies of transcription-coupled nucleotide excision repair and ALKBH3-mediated alkylation repair. It can be employed to investigate R-loop accumulation by DNA/RNA immunoprecipitation, assess DNA damage responses through ??H2AX immunofluorescence and comet assays, and analyze transcription recovery using nascent RNA RT-qPCR or RNA-seq. Moreover, these cells provide a platform for drug sensitivity profiling with alkylating agents such as MMS and cisplatin, and for exploring the molecular basis of diseases linked to ASCC3, including autosomal recessive intellectual disability, microcephaly, Barrett??s esophagus, and esophageal adenocarcinoma. For further details and customization options, please contact Ascent Research.
