ICK Knockout HAP1 Cell Line

The choice depends on whether you are studying ICK (intestinal cell kinase, CILK1)'s role as a ciliary kinase or modeling endocrine-cerebro-osteodysplasia (ECO) syndrome and other ICK-related ciliopathies. The Knockout line is the standard tool for asking whether ICK is required for these processes — ICK and its paralog MAK constitute the RCK family of CDK-related kinases that regulate ciliary length and microtubule dynamics; ICK phosphorylates KIF3A and other ciliary substrates to control intraflagellar transport. Overexpression is useful for studying ICK in heterologous expression contexts. For ciliary biology research, the EDITGENE ICK Knockout in HAP1 enables study of CILK1/ICK biology. MAK paralog (Gene ID 4117, also available as EDITGENE MAK Knockout in HAP1) expression analysis aids interpretation given functional overlap in the RCK family. ICK biallelic loss-of-function mutations cause endocrine-cerebro-osteodysplasia syndrome (ECO, lethal multi-system ciliopathy) and juvenile myoclonic epilepsy — disease variant rescue enables genotype-function studies. Rescue with wild-type or kinase-dead ICK is the standard specificity control.

Primary applications: • Ciliary length regulation: cilium length and frequency analysis given ICK's role in cilium control. • KIF3A phosphorylation: phospho-KIF3A Western blot analysis to characterize ICK-dependent IFT regulation. • ECO syndrome modeling: rescue with patient-derived ICK mutations for genotype-function studies of endocrine-cerebro-osteodysplasia. • RCK family comparative studies: parallel analysis with MAK Knockout in HAP1 (also available) for paralog-specific functional dissection. EDITGENE recommends this model for researchers investigating ICK/CILK1 kinase biology, ciliopathy mechanisms, and RCK family functional specialization.

Yes. ICK rescue experiments require attention to ciliary kinase architecture: • Construct design: use a codon-modified ICK sequence with a small C-terminal tag (FLAG, HA). ICK has N-terminal kinase domain with characteristic CDK-like architecture and C-terminal regulatory region — preserve all elements. • Kinase-dead rescue: K33A or D146A mutations in the catalytic kinase domain abolish catalytic activity and serve as the standard specificity control. • ECO syndrome mutation rescue: patient-derived ICK mutations enable disease genotype-function studies. • Functional readout: rescue should restore ciliary length regulation and KIF3A phosphorylation. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.