The ATP11C Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line, designed to disrupt ATP11C, a P4-type ATPase flippase. This heterogeneous knockout model, created via CRISPR-mediated gene disruption, enables loss-of-function studies of ATP11C in a pooled format, circumventing clonal variability. The product serves as a versatile system for probing phospholipid asymmetry, apoptosis, and immune evasion in colorectal cancer.
The HT29 cell line originates from a colorectal adenocarcinoma in a 44-year-old female and is widely employed to model intestinal epithelial biology. These cells are mucus-secreting and retain epithelial characteristics, with mutations in p53 and APC that drive key oncogenic pathways. As a well-established cancer model, HT29 provides a relevant platform to investigate how ATP11C loss influences malignant epithelial phenotypes such as altered membrane dynamics and drug resistance.
ATP11C constitutes the catalytic subunit of a flippase complex with TMEM30A (CDC50A), which actively internalizes phosphatidylserine (PS) at the plasma membrane, preserving lipid asymmetry and preventing external PS exposure. Its activity is regulated by Ca2+, PKC phosphorylation, and caspase-3-mediated cleavage, while transcription is controlled by IKZF1. In B lymphocytes, ATP11C is critical for pre-BCR signaling, functioning downstream of pre-BCR and upstream of SLP-65 (BLNK), BTK, PLC??2, and Ca2+ flux to drive B-cell development and integrin LFA-1 activation. In epithelial cells, ATP11C modulates membrane architecture and apoptosis sensitivity.
In HT29 colorectal adenocarcinoma cells, disruption of ATP11C is expected to perturb PS asymmetry, potentially leading to aberrant externalization of PS and altered immune recognition. This model facilitates the study of flippase-mediated apoptosis resistance, a process often exploited by cancer cells to evade phagocytic clearance. Moreover, it enables investigation of ATP11C??s contribution to epithelial membrane organization, polarity, and barrier function in a mucinous colorectal cancer context.
Researchers can utilize this polyclonal knockout population for diverse assays, such as Annexin V flow cytometry to quantify PS exposure, NBD-PS flippase activity measurements, and immunoblotting or RT-qPCR for ATP11C expression. Functional analyses include apoptosis and viability assays (MTT), colony formation, and immunofluorescence for flippase localization. Co-immunoprecipitation with TMEM30A and Ca2+ flux experiments aid in network dissection. Furthermore, co-culture systems with lymphoid cells allow exploration of ATP11C??s role in immune signaling, supporting applications in cancer biology, immunology, and drug screening. For additional details, contact Ascent Research.
