The ATF7IP Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 haploid human cell line. This loss-of-function model has been engineered through CRISPR/Cas9-mediated gene disruption of ATF7IP, resulting in a heterogeneous pool of cells carrying diverse indel mutations at the target locus. The polyclonal format provides a convenient and robust system for studying ATF7IP function without the need for single-cell clonal isolation, enabling interrogation of gene function in a near-haploid genetic background that simplifies genotype-phenotype analyses.
The parental HAP1 cell line is a near-haploid human fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) cell line, which was isolated from a male patient. HAP1 cells retain a haploid karyotype for most chromosomes, except for a disomic region of chromosome 8 and a portion of chromosome 15, making them an ideal host for CRISPR-based functional genomics and haploid genetic screening. Their leukemic origin also renders them particularly relevant for studying myeloid neoplasms and leukemia-associated pathways, including epigenetic regulators such as ATF7IP.
ATF7IP (also known as MCAF1) functions as a transcriptional coregulator that bridges stress-activated transcription factors with the epigenetic silencing machinery. It directly interacts with ATF7, a transcription factor activated by MAP kinases p38 and JNK in response to cellular stress. Upon recruitment, ATF7IP assembles a repressor complex containing MBD1 and the histone methyltransferase SETDB1, which catalyzes trimethylation of histone H3 at lysine 9 (H3K9me3). Interactions with heterochromatin protein 1 (HP1/CBX5) and the transcription factor SP1 lead to heterochromatin formation and stable gene silencing. Through these interactions, ATF7IP couples stress signaling pathways to heritable chromatin modifications and DNA methylation-mediated transcriptional repression.
In the HAP1 haploid background, disruption of ATF7IP offers a unique window into the immediate consequences of losing ATF7IP-mediated epigenetic silencing. Since HAP1 cells possess only one functional allele, knockout effects are unmasked without compensation from a second allele, providing a sensitized background for identifying targets of ATF7IP-dependent regulation. This model is particularly relevant for investigating aberrant epigenetic silencing in leukemia and other myeloid malignancies, where ATF7IP has been implicated in shaping the chromatin landscape and gene expression programs that drive disease progression.
Researchers can leverage the ATF7IP Knockout HAP1 Polyclonal Cells in a broad range of assays, including Western blotting and RT-qPCR to confirm protein loss and transcript changes, ChIP-qPCR to assess histone modifications and transcription factor binding, co-immunoprecipitation to probe protein interactions, and RNA-seq or bisulfite sequencing for genome-wide expression and DNA methylation profiling. These cells are suited for functional genomics screens, drug sensitivity assays to evaluate therapeutic responses, and validation of ATF7IP as a drug target in leukemia. For technical inquiries or ordering information, please contact Ascent Research.
