LRP6 Knockout HAP1 Cell Line

The choice depends on whether you are studying LRP6 (low-density lipoprotein receptor-related protein 6)'s role as the principal Wnt co-receptor or modeling its associations with cardiovascular disease and metabolic phenotypes. The Knockout line is the standard tool for asking whether LRP6 is required for canonical Wnt/β-catenin signaling — LRP6 partners with Frizzled receptors as the obligate co-receptor for canonical Wnt ligands, with intracellular PPPSP motifs that become phosphorylated upon Wnt activation to recruit Axin and disassemble the β-catenin destruction complex. Overexpression is useful for studying LRP6 gain-of-function effects. For Wnt signaling research, the EDITGENE LRP6 Knockout in HAP1 enables study of canonical Wnt signaling. LRP5 paralog expression analysis aids interpretation given functional overlap as Wnt co-receptors. Rescue with wild-type or PPPSP-motif-mutant LRP6 enables structure-function studies. The knockout is valuable for testing LRP6 modulators (anti-LRP6 antibodies in oncology development) and studying LRP6-mutation-associated coronary artery disease (R611C variant) genotype-function relationships.

Primary applications: • Canonical Wnt/β-catenin signaling: TCF/LEF reporter assays, β-catenin stabilization, and Wnt target gene (AXIN2, LGR5, MYC) expression following Wnt3a or Wnt1 stimulation in LRP6-null cells. • Wnt co-receptor studies: LRP5 expression analysis to interpret LRP6-specific functions. • Phospho-LRP6 analysis: phospho-LRP6 (PPPSP motif) Western blot following Wnt stimulation to characterize receptor activation. • Cardiovascular disease modeling: rescue with R611C and other CAD-associated LRP6 variants for genotype-function studies of premature coronary artery disease. EDITGENE recommends this model for researchers investigating canonical Wnt signaling, LRP6 co-receptor biology, and cardiovascular disease genetics.

Yes. LRP6 rescue experiments are well-established for Wnt signaling research: • Construct design: use a codon-modified LRP6 sequence with a small intracellular C-terminal tag (FLAG, HA). LRP6 has extracellular β-propeller-EGF repeats, single transmembrane span, and intracellular cytoplasmic tail with PPPSP motifs — preserve all elements. • PPPSP-mutant rescue: PPPSP motif phosphorylation site mutations abolish Wnt signaling activation and serve as the standard specificity control. • CAD variant rescue: R611C and other coronary artery disease-associated LRP6 variants enable disease genotype-function studies. • Functional readout: rescue should restore Wnt-induced phospho-LRP6, β-catenin stabilization, and TCF/LEF reporter activity. 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.