The HCFC1R1 Knockout HAP1 Polyclonal Cells product provides a carefully engineered pool of CRISPR/Cas9-edited HAP1 cells in which the HCFC1R1 gene has been disrupted. This polyclonal population consists of a heterogeneous mix of knockout genotypes generated by CRISPR/Cas9-mediated gene targeting, offering a robust loss-of-function model without the need for single-cell cloning. The cell mixture is designed for high-throughput genetic screens and functional genomics applications, enabling the study of gene disruptions in a physiologically relevant context. As a polyclonal knockout product, it retains cellular diversity while ensuring effective abrogation of HCFC1R1 protein expression across the population.
The host cell line, HAP1, is a widely utilized near-haploid human fibroblast line derived from the KBM-7 chronic myeloid leukemia isolate. Its near-haploid karyotype simplifies genetic analysis by eliminating the complexity of diploid gene dosage, making it an ideal platform for knockout and genetic screening studies. HAP1 cells maintain key features of mesenchymal cells and exhibit stable growth characteristics, facilitating reproducible experimental outcomes. The line??s origin from a hematological malignancy background further positions it as a relevant model for exploring cancer-associated pathways, particularly those linked to chromatin biology and transcriptional control.
HCFC1R1 (also known as ZNF143) encodes a zinc finger transcription factor that functions as a key insulator protein, binding to SphI consensus motifs found in numerous gene promoters. It facilitates chromatin looping and communication between distal regulatory elements and core promoters, thereby mediating finely tuned transcriptional programs. Upstream, HCFC1R1 expression is regulated in a cell cycle-dependent manner. Downstream, it targets SphI element-containing promoters and interacts directly with the transcription machinery, including RNA polymerase II complexes, as well as chromatin remodeling factors. Through these interactions, HCFC1R1 orchestrates transcriptional activation or repression of target genes, influencing processes such as cell cycle progression and chromatin organization.
In the HAP1 background, knockout of HCFC1R1 disrupts insulator function and deregulates the transcriptional programs normally coordinated by this factor. The loss of ZNF143 binding at insulator sites impairs proper chromatin looping, leading to aberrant expression of downstream target genes. Given HAP1??s origin from a chronic myeloid leukemia line, the model holds particular significance for studying hematological malignancies and cancer, where dysregulation of transcriptional and cell cycle control is common. Researchers can use this system to dissect the consequences of insulator protein deficiency in a simplified genetic environment, probing the interplay between chromatin architecture and gene expression.
These polyclonal knockout cells are suited for a range of experimental assays, including chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) to examine transcription factor binding, RNA sequencing to assess transcriptome-wide changes, western blotting for protein-level confirmation, luciferase reporter assays to test promoter activity, and immunofluorescence for subcellular localization studies. The model supports functional genomics screens, gene regulation analysis, and disease-relevant investigations into transcriptional control mechanisms. For technical inquiries, contact Ascent Research.
