The ATP2A1 Knockout HeLa Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ATP2A1 gene. This product provides a genetically disrupted ATP2A1 allele pool in the widely used HeLa background, eliminating the need for single-clone isolation and enabling functional analyses within a heterogeneous editing landscape. The polyclonal format facilitates robust assessment of ATP2A1-dependent phenotypes while minimizing clonal artifacts, making it suitable for high-content screening and population-level assays.
The host HeLa cell line is an immortalized human cervical adenocarcinoma epithelial model positive for HPV18, with well-characterized inactivation of the p53 and Rb tumor suppressors by viral E6 and E7 oncoproteins. These cells exhibit a highly aneuploid karyotype and uncontrolled proliferation, providing a reproducible and experimentally tractable system for cancer biology and calcium signaling studies. Their robust growth and ease of transfection further support CRISPR-based genome editing and downstream functional assays.
ATP2A1 encodes the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA1), a primary pump responsible for cytosolic calcium reuptake into the ER lumen, essential for calcium homeostasis and muscle contraction. Its activity is modulated by upstream regulators including thyroid hormone (T3), MyoD/MEF2 transcription factors, sarcolipin (SLN), and the DWORF peptide. SERCA1 interacts directly with SLN, DWORF, myoregulin, and calmodulin, and its function impacts downstream effectors such as calmodulin, CaMKII, calcineurin, and NFAT. Loss of SERCA1 elevates cytosolic Ca2+ levels, triggering ER stress and unfolded protein responses (UPR) via BiP and CHOP, which can lead to caspase-12-mediated apoptosis or calcium-dependent transcriptional reprogramming through NFAT and CREB.
In the HeLa background, ATP2A1 knockout disrupts the already delicately balanced calcium signaling networks that influence cancer cell proliferation, survival, and stress adaptation. The resultant chronic ER calcium depletion and sustained cytosolic calcium overload activate UPR sensors and may shift cell fate toward apoptosis or adaptive survival, providing a unique model to dissect calcium-mediated oncogenic mechanisms and tumor cell vulnerabilities. This system is particularly valuable for examining cross-talk between ER stress and HPV-driven transformation, as well as for interrogating the role of SERCA1 in cancer cell resilience against proteotoxic insults.
Researchers can employ these polyclonal knockout cells in a wide range of assays to explore calcium-dependent signaling in cancer, evaluate ER stress/UPR pathways, screen for SERCA modulators, and investigate apoptosis regulation. Typical workflows include intracellular calcium imaging with Fluo-4 AM, Western blotting for SERCA1 and UPR markers, RT-qPCR for calcium-handling gene expression, flow cytometric annexin V/PI apoptosis assays, cell viability measurements, co-immunoprecipitation of SERCA1 interactors, and transcriptomic profiling by RNA-seq. Drug sensitivity testing can identify compounds that synergize with SERCA1 loss. For further information on availability and experimental support, please contact Ascent Research.
