The DNMT3L Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human HAP1 cell line, offering a targeted loss-of-function model for the DNMT3L gene. This product provides a genetically diverse pool of cells harboring disruptions in DNMT3L, enabling the study of gene function without the need for clonal isolation. The polyclonal format is particularly suited for applications requiring population-level analyses of DNA methylation dynamics and epigenetic regulation.
HAP1 is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line, which carries the BCR-ABL fusion oncogene. Its haploid karyotype facilitates straightforward genetic manipulation and phenotype analysis, as single-allele disruptions can unmask recessive phenotypes. This feature makes HAP1 a powerful host for CRISPR-based screens and functional genomics, especially in the context of leukemia biology and cancer epigenetics.
DNMT3L functions as a catalytically inactive DNA methyltransferase cofactor that forms complexes with the de novo methyltransferases DNMT3A and DNMT3B. Through its recognition of unmethylated histone H3K4 tails, DNMT3L stimulates the methylation of imprinting control regions and transposable elements, including LINEs and SINEs, thereby directing epigenetic silencing. It interacts with heterochromatin protein 1 (HP1) and is recruited to chromatin via histone H3 modifications. Upstream regulation involves retinoic acid and transcription factors SOX9 and DMRT1, as well as EZH2-mediated histone methylation. Downstream targets include imprinted loci such as H19 and IGF2, whose methylation status depends on DNMT3L activity. The pathway integrates S-adenosylmethionine as the methyl donor, and involves methyl-CpG-binding domain proteins (MBDs) and histone deacetylases (HDACs) in the maintenance of silencing.
In the HAP1 leukemia-derived context, DNMT3L knockout provides a unique platform to investigate de novo methylation mechanisms in cancer cells, where aberrant DNA methylation is a hallmark. Loss of DNMT3L disrupts proper imprinting and transposon silencing, potentially contributing to genomic instability and oncogenic transformation. The near-haploid nature of HAP1 allows for high-efficiency screening to identify genetic interactions and dependencies related to DNMT3L function, offering insights into imprinting disorders such as Beckwith-Wiedemann syndrome and Silver-Russell syndrome, as well as infertility.
This polyclonal knockout cell model is designed for a wide range of epigenetic research applications, including the study of de novo DNA methylation, genomic imprinting, transposon silencing, and epigenetic gene regulation. Researchers can employ bisulfite sequencing, methylation-specific PCR, methylated DNA immunoprecipitation (MeDIP), and ChIP-qPCR for DNMT3A/B to assess methylation changes. Additionally, RNA-seq, western blot, immunofluorescence, and transposon expression analysis can elucidate downstream molecular effects. The polyclonal population is also amenable to haploid genetic screens for synthetic lethality or pathway modulators. For further information or custom requests, please contact Ascent Research.
