The ACVR2A Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited heterogeneous cell pool derived from the human near-haploid HAP1 line, featuring targeted disruption of the ACVR2A gene. This polyclonal population avoids clonal artifacts and provides a robust loss-of-function model for functional studies and signaling assays. The knockout cells are supplied as a mixed population, enabling experimental consistency across replicate batches and supporting bulk analyses such as pooled screening. By circumventing clonal selection, it reduces the risk of isolated off-target effects and ensures a representative loss-of-function phenotype.
The parental HAP1 cell line is a near-haploid human chronic myeloid leukemia cell line (45,X) derived from KBM-7, widely used for CRISPR-based genetic screens because its haploid genome simplifies gene-editing and eliminates functional redundancy. HAP1 cells retain core hematopoietic signaling pathways, offering an ideal background for studying ACVR2A-related mechanisms in cancer and cell biology. The presence of a single allele at most loci ensures that CRISPR-induced mutations lead to loss of gene function, making it a stringent system for genotype-phenotype correlation.
ACVR2A is a type II receptor for Activin A, Activin B, Myostatin, and GDF11. Ligand binding induces recruitment and phosphorylation of type I receptors ACVR1B (ALK4) or ACVR1C (ALK7), which activate the SMAD2/3?CSMAD4 transcriptional complex to regulate genes such as SERPINE1 and CCN2. Additionally, ACVR2A triggers non-canonical MAPK/ERK and PI3K/AKT signaling through MAPK1 and AKT1. Interactors including FKBP1A and ZFYVE9 modulate receptor output. This dual signaling mode allows ACVR2A to finely tune cellular outcomes depending on context. Disruption of ACVR2A thus impairs both canonical and non-canonical branches, enabling comprehensive dissection of ligand-specific responses.
ACVR2A mutations are prevalent in microsatellite instability-high colorectal cancer, gastric and endometrial cancers, and pituitary adenomas, often leading to impaired tumor-suppressive activin/TGF-?? signaling. In the HAP1 near-haploid system, ACVR2A disruption generates functional hemizygosity, revealing robust phenotypes suitable for dissecting oncogenic dependencies and therapeutic vulnerabilities. This model thus enables the examination of pathway alterations that drive carcinogenesis and influence drug sensitivity.
Applications include western blot for phospho-SMAD2, RT-qPCR of downstream effectors, RNA-seq, and immunofluorescence for SMAD nuclear shuttling. Functional assays such as proliferation, apoptosis flow cytometry, phospho-ELISA, and CAGA-luciferase reporters allow quantitative pathway analysis, supporting target validation, drug testing, and genetic screens. These approaches facilitate detailed mechanistic studies and high-throughput screening campaigns. For additional information, contact Ascent Research.
