This product is a CRISPR/Cas9-edited polyclonal knockout cell population of HCT 116, a human colorectal carcinoma epithelial cell line. The polyclonal nature yields a mixed pool of edited cells, enabling loss-of-function studies of ACVR2A in a bulk culture format. Targeted disruption of the ACVR2A gene was achieved using CRISPR/Cas9, resulting in a population with diverse editing outcomes, appropriate for assays that assess average gene knockout effects at the population level.
The HCT 116 parental line is a widely utilized colorectal cancer model characterized by an epithelial morphology and key genetic alterations: an activating KRAS G13D mutation, CTNNB1 S45 deletion, microsatellite instability-high (MSI-H) status, and wild-type p53. These features drive constitutive Wnt/beta-catenin and MAPK/ERK pathway activation and defective DNA mismatch repair, making the line ideal for studying tumor suppressor mechanisms and oncogenic signaling crosstalk.
ACVR2A encodes the type II receptor for activin ligands, including activin A, activin B, inhibin, and nodal. Upon ligand binding, ACVR2A associates with type I receptors such as ACVR1B (ALK4) or ACVR1C (ALK7), leading to phosphorylation of SMAD2 and SMAD3. Phosphorylated SMAD2/3 complex with SMAD4 and translocate to the nucleus to regulate transcription of target genes like CDKN1A (p21) and c-MYC. Regulatory interactions with FKBP12, SMURF1, SMURF2, and ZFYVE9 (SARA) fine-tune the signaling output, ultimately controlling cell proliferation, differentiation, and apoptosis.
In the HCT 116 context, ACVR2A disruption is especially relevant because MSI-H colorectal cancers frequently harbor ACVR2A mutations, contributing to tumor progression by impairing growth-inhibitory and pro-apoptotic signaling. Combined with the constitutive KRAS and Wnt pathway activation in HCT 116 cells, loss of ACVR2A provides a powerful system to dissect the interplay between activin/SMAD signaling and other oncogenic pathways, offering insights into MSI-H tumor biology and potential therapeutic targets.
These polyclonal knockout cells are suitable for a range of functional analyses, including phospho-SMAD2/3 Western blotting, RT-qPCR for downstream targets, and phenotypic assays such as MTT or BrdU proliferation and Annexin V apoptosis measurements. Colony formation, xenograft tumor growth studies, and CAGA-luciferase reporter assays can further define the role of ACVR2A in tumorigenesis. The model also supports drug screening for TGF-beta pathway modulators. For technical assistance, please contact Ascent Research.
