KITLG Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the KITLG gene in the HAP1 human near-haploid cell line. This loss-of-function model enables the study of KITLG-dependent signaling in a hematopoietic progenitor-like cancer cell background. The polyclonal format captures a heterogeneous pool of edited alleles, offering a robust system for evaluating gene function without clonal bias. The knockout is introduced via CRISPR/Cas9-mediated gene disruption, eliminating functional KITLG protein expression, and is suitable for a wide range of downstream functional assays.
The HAP1 host cell line is a near-haploid, adherent, fibroblast-like cell line derived from the male KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype simplifies genetic manipulation and facilitates unambiguous genotype-phenotype correlations. HAP1 cells retain characteristics of hematopoietic progenitor cells, making them a valuable model for studying hematopoiesis, leukemia biology, and signal transduction. The cells grow as a monolayer, are easy to transfect, and are widely used in functional genomics and drug screening applications.
KITLG encodes stem cell factor (SCF), the ligand for the c-KIT receptor tyrosine kinase. Upon binding, KITLG induces receptor dimerization and autophosphorylation, activating multiple downstream cascades including PI3K/AKT, RAS/MAPK, and JAK/STAT. Key upstream regulators include HIF1A, NF-??B, SOX10, and TGFB1, which modulate KITLG expression under various physiological and pathological conditions. Downstream effectors such as AKT, ERK1/2, STAT3, BCL2, and CCND1 mediate survival, proliferation, and differentiation signals. KITLG signaling involves adaptor proteins GRB2, SHC1, and SOS1, which couple the activated receptor to RAS-MAPK activation. This pathway is critical for the maintenance and migration of hematopoietic stem cells, melanocytes, and germ cells.
In the HAP1 hematopoietic progenitor-like background, KITLG knockout disrupts autocrine or paracrine SCF/c-KIT signaling, providing a physiologically relevant context to investigate the role of this axis in leukemia and other KIT-driven malignancies. Given the near-haploid nature of HAP1 cells, the knockout model allows clean dissection of KITLG contributions without genetic redundancy from a diploid genome. This model is particularly useful for studying signaling mechanisms underlying mastocytosis, gastrointestinal stromal tumors, and acute myeloid leukemia, where KIT mutations are prevalent. Additionally, the model can be used to explore resistance mechanisms to KIT-targeted therapies.
Researchers can employ this knockout model in a variety of experimental workflows: assessing KITLG-dependent activation of downstream kinases by western blotting for phospho-KIT, AKT, and ERK1/2; quantifying KITLG transcript levels by RT-qPCR; measuring KIT surface expression by flow cytometry; and evaluating functional consequences such as proliferation, migration, and drug sensitivity. The polyclonal population is ideal for pooled functional genomics screens and for generating reproducible data across multiple assays. For additional information, please contact Ascent Research.
