The DSCC1 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population that provides a loss-of-function model for DSCC1 (DNA replication and sister chromatid cohesion 1) in a human haploid cell line. By disrupting the DSCC1 gene, these polyclonal cells facilitate investigation of the gene??s essential roles in DNA replication and chromosome cohesion without requiring isolation of single clones. The polyclonal format captures a spectrum of editing outcomes, enabling robust study of DSCC1-dependent processes.
The host cell line, HAP1, is a near-haploid human cell line derived from a patient with chronic myelogenous leukemia (CML). Its haploid karyotype simplifies gene knockout studies, as only one allele requires disruption, and it has become a widely adopted model for functional genomics and genetic screening. HAP1 cells retain fundamental signaling and replication pathways, making them suitable for dissecting the molecular mechanisms of DSCC1 in a genetically clean background.
DSCC1 encodes a core subunit of the CTF18-RFC alternative replication factor C complex, which is essential for loading proliferating cell nuclear antigen (PCNA) onto chromatin during S phase. This PCNA loading function is a prerequisite for the establishment of sister chromatid cohesion, a process that also requires the recruitment of the cohesin complex, including subunits SMC1A, SMC3, RAD21, and STAG1/2. DSCC1 expression is regulated by E2F transcription factors in a cell cycle-dependent manner. Within the CTF18-RFC complex, DSCC1 interacts with CTF18, CTF8, and RFC2?C5 to facilitate PCNA recruitment. Disruption of DSCC1 abrogates this pathway, leading to defective cohesion, replication fork instability, and increased genome instability.
In the HAP1 background, knockout of DSCC1 is predicted to cause pronounced sister chromatid cohesion defects and replication stress, owing to the haploid state??s inability to buffer the loss of this critical factor. This model is particularly relevant for investigating DSCC1-related neurodevelopmental disorder, where recessive mutations cause severe developmental phenotypes, and for studying the role of cohesion dysfunction in cancer. The polyclonal cell population allows researchers to observe phenotypic heterogeneity and to perform experiments that average across multiple disruptive edits, mimicking population-level effects of DSCC1 deficiency.
These polyclonal knockout cells are well-suited for a variety of downstream applications, including chromosome spread assays to assess cohesion defects, immunofluorescence microscopy to examine cohesin complex localization, and DNA fiber assays to measure replication fork dynamics. Additionally, flow cytometry enables cell cycle profiling, while western blotting can quantify PCNA loading and cohesin subunit levels. The model supports viability and apoptosis assays under replication stress conditions and is an excellent platform for cohesin inhibitor drug screening and functional genomics studies of the cohesion pathway. For further technical inquiries, please contact Ascent Research.
