The CCIN Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout population in the HAP1 cell line with targeted disruption of the CCIN gene. This pool enables loss-of-function studies of calicin in a near-haploid human context. The polyclonal format maintains genetic diversity while abrogating calicin expression, making it suitable for high-throughput screening and pooled functional genomics. Researchers can investigate calicin-dependent processes without wild-type interference.
HAP1 cells derive from the KBM-7 CML line with a near-haploid karyotype, simplifying functional genomics and screening. They retain BCR-ABL signaling dependence, serving as a hematopoietic cancer model for drug sensitivity studies. The reduced gene redundancy facilitates knockout phenotype interpretation, though CCIN is not endogenously expressed, requiring ectopic expression for functional studies.
Calicin is a structural component of the sperm perinuclear theca, participating in sperm head shaping and acrosome integrity. It interacts with cylicin-1, cylicin-2, actin, and calmodulin to reorganize the actin cytoskeleton and assemble the perinuclear theca. Transcriptionally regulated by CREM, SOX5, and SPZ1, calicin functions downstream of spermatogenic signals. While these networks are well-defined in germ cells, their roles in other cell types are unclear. The CCIN knockout HAP1 model allows dissection of these interactions when calicin is ectopically expressed, enabling analysis of its protein interaction network.
While calicin’s primary function is in male fertility, its knockout in HAP1 cells provides a platform for studying perinuclear theca proteins in a non-germline setting. The near-haploid genome reduces compensatory effects and improves gene editing efficiency, enabling clean assessment of calicin’s impact on actin dynamics. Coupled with ectopic expression, the model helps elucidate calicin’s role in cytoskeletal organization and may inform mechanisms of spermatogenic failure. It also supports drug screening for compounds that modulate actin-calicin interactions.
Applications include functional genomics, co-immunoprecipitation-based interaction studies, and male infertility research with ectopic CCIN. Knockout validation uses western blotting, RT-qPCR, and immunofluorescence, while functional assays involve actin staining, viability, and drug sensitivity tests. The polyclonal pool is advantageous for pooled CRISPR screens and interaction proteomics. For inquiries, contact Ascent Research.
