The BACE2 Knockout HCT 116 Polyclonal Cells constitute a CRISPR/Cas9?edited polyclonal knockout population of the human colorectal carcinoma HCT 116 cell line, featuring targeted disruption of the BACE2 gene. This loss?of?function model eliminates BACE2 ???secretase activity, enabling investigation of its proteolytic substrates and downstream pathways without clonal selection bias.
HCT 116 is a widely used epithelial colon carcinoma line harboring KRAS G13D and PIK3CA activating mutations, along with MLH1 deficiency that confers microsatellite instability?high (MSI?H). These genetic features make it a well?characterized model for colorectal cancer signaling, drug responses, and DNA mismatch repair studies. The cell line??s rapid growth and defined tumorigenic properties facilitate high?throughput functional genomics and drug screening.
BACE2 encodes a transmembrane aspartyl protease that preferentially cleaves type I membrane substrates at ???secretase sites, including APP, Notch1, Notch2, TMEM27, and PMEL. Its transcription is regulated by PPAR??, various transcription factors, and microRNAs, while its activity and subcellular localization are influenced by interactions with the ???secretase complex and reticulon proteins. Proteolytic processing of APP generates A?? peptides that aggregate in Alzheimer??s disease; shedding of TMEM27 controls ???cell proliferation and insulin secretion; and PMEL cleavage is essential for amyloid fibril formation within melanosomes. Consequently, BACE2 occupies a pivotal node connecting neurodegeneration, Notch signaling, melanogenesis, and metabolic regulation.
In HCT 116 colorectal carcinoma cells, BACE2 knockout eliminates ???secretase activity toward Notch and APP, abrogating NICD release and A?? production. Because Notch signaling drives colorectal cancer stem cell maintenance, epithelial?Cmesenchymal transition, and chemoresistance, loss of BACE2?mediated Notch processing can reveal context?dependent vulnerabilities. Additionally, the MSI?H/MLH1?deficient genomic background permits investigation of BACE2??s interplay with DNA mismatch repair and the response to genotoxic agents such as 5?fluorouracil. This knockout model thus provides a unique platform to study tumor?intrinsic protease functions and identify novel biomarkers or therapeutic targets.
Research applications include Western blotting to confirm BACE2 depletion, RT?qPCR for mRNA quantification, Notch reporter and A?? ELISA assays to monitor substrate processing, and proteomics for global substrate profiling. Functional studies such as migration/invasion and apoptosis assays can dissect BACE2??s role in tumor progression. This polyclonal population supports projects ranging from cancer biology and colorectal carcinoma drug screening to Alzheimer??s disease and diabetes research. For further details, please contact Ascent Research.
