The L1TD1 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population in the HAP1 human near-haploid cell line, targeting the L1TD1 gene. This heterogeneous pool of edited alleles offers a robust loss-of-function model, circumventing clonal artifacts. The CRISPR system disrupts L1TD1 without predefined editing patterns, providing an unbiased tool for functional studies in a haploid genetic environment tailored for screening, functional genomics, and pathway interrogation.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line through near-haploidization, retaining a haploid karyotype except for a disomic chromosome 15. This simplicity facilitates unambiguous gene-disruption phenotypes, making it a preferred model for haploid genetics and cancer research. The adherent cells maintain hematopoietic features and are extensively used in CRISPR-based studies for target validation, signaling dissection, and oncology.
L1TD1 is an RNA-binding protein controlled by core pluripotency transcription factors NANOG, POU5F1 (OCT4), and SOX2, and is essential for embryonic stem cell self-renewal and pluripotency. It binds LINE-1 ORF1p and modulates retrotransposon activity, safeguarding genomic integrity. L1TD1 functions within a regulatory network including KLF4 and MYC, and its expression is influenced by Wnt and TGF-beta pathways. Disruption of L1TD1 perturbs the pluripotency gene network, linking retrotransposon regulation to stem cell identity.
In the HAP1 leukemic context, L1TD1 knockout allows dissection of pluripotency-related pathways in cancer stem cell biology. L1TD1 is implicated in germ cell tumors and cancer stem cell malignancies, and its loss in a near-haploid system reveals un-compensated signaling effects. Researchers can explore how NANOG, POU5F1, and SOX2-driven programs contribute to oncogenic dedifferentiation when L1TD1 is disrupted, providing a platform to study tumor-initiating cells.
The knockout pool supports diverse applications, including pluripotency maintenance studies, LINE-1 retrotransposon regulation, and cancer stem cell research. Assays such as Western blotting, RT-qPCR, RNA-seq, immunofluorescence, flow cytometry, and reporter assays are routinely applied. Co-immunoprecipitation maps interactions with NANOG, POU5F1, and ORF1p, while apoptosis and migration/invasion assays assess phenotypic outcomes. For further information and tailored experimental support, please contact Ascent Research.
