The KIAA1671 Knockout HAP1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the KIAA1671 gene has been disrupted. This pooled cell format offers a versatile loss-of-function model for studying centrosomal protein function and ciliogenesis without the selection of a single clonal isolate.
These knockout cells are engineered in the HAP1 cell line, a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) cell line originally isolated from a male patient. The HAP1 adherent cells contain a single copy of most chromosomes except chromosome 8, facilitating efficient gene disruption and functional genetic studies. Their haploid karyotype makes them an ideal platform for generating knockouts and screening for phenotypes with reduced genetic redundancy.
The KIAA1671 gene encodes a centriolar satellite protein critical for proper cilium assembly and maintenance. The protein functions within the centrosome cycle and ciliogenesis pathway, acting downstream of RFX transcription factors and cell cycle regulators to control microtubule organization and ciliary membrane trafficking. KIAA1671 forms complexes with key centriolar satellite components including PCM1, CEP290, and CCDC66, and its loss disrupts the transport and localization of these factors. Consequently, KIAA1671 knockout impairs Hedgehog signaling by affecting the processing and activity of GLI transcription factors, as the primary cilium serves as a signaling hub for the Hedgehog pathway. Representative pathway components affected include IFT88, SMO, and GLI1, linking KIAA1671 to ciliary trafficking and signal transduction.
In the HAP1 near-haploid background, disruption of KIAA1671 yields a clean genetic model to dissect ciliogenesis mechanisms with minimal confounding from a second functional allele. The polyclonal nature of this knockout population provides a heterogeneous pool of edited cells, enabling robust phenotypic analyses while avoiding clonal artifacts. This model is particularly valuable for studying centrosome-related disorders and ciliopathies, as HAP1 cells retain functional ciliary assembly pathways and respond to Hedgehog pathway modulators.
Typical applications of these KIAA1671 knockout polyclonal cells include immunofluorescence staining for ciliary markers such as acetylated tubulin and ARL13B to assess cilia formation, western blotting for GLI proteins to monitor Hedgehog pathway activity, and RT-qPCR of Hedgehog target genes to quantify transcriptional output. The cells can be used for drug screening in ciliopathy research, functional genomics studies leveraging the haploid genome, and investigation of centrosomal protein trafficking. Genotyping by PCR and Sanger sequencing can confirm KIAA1671 disruption at the population level. For additional information, researchers are encouraged to contact Ascent Research.
