DYNC2LI1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the targeted disruption of the human DYNC2LI1 gene. This knockout model provides a powerful loss-of-function tool to investigate the roles of the light intermediate chain 1 of cytoplasmic dynein-2 in primary cilia biology and associated signaling cascades.
Originating from the HeLa cell line, a human cervical adenocarcinoma epithelial line derived from Henrietta Lacks, these cells are characterized by HPV18 positivity and expression of the viral oncoproteins E6 and E7, which respectively inhibit p53 and Rb, conferring an immortalized phenotype suitable for extensive experimental manipulation. The epithelial background and widespread use of HeLa make them an ideal host for studying ciliary dynamics and cancer-relevant pathways.
DYNC2LI1 encodes a subunit of the cytoplasmic dynein-2 retrograde motor complex, critical for intraflagellar transport (IFT) in primary cilia. It interacts with the heavy chain DYNC2H1 and intermediate chains WDR34 and WDR60 to facilitate trafficking of IFT-A/B complexes from the ciliary tip to the base. This retrograde movement is essential for ciliogenesis and hedgehog signaling. Upstream, RFX3, FOXJ1, and serum deprivation regulate DYNC2LI1 expression. Downstream, functional dynein-2 processes GLI2 and GLI3 transcription factors, thereby modulating hedgehog target genes such as PTCH1, GLI1, and CCND1. Disruption of DYNC2LI1 results in accumulation of proteins like KIF7 and SUFU at the ciliary tip, impairing hedgehog and cilia-dependent TGF-?? signaling.
When cultured under serum deprivation, HeLa cells form primary cilia, making them a relevant model for DYNC2LI1 knockout studies. Loss of this gene recapitulates molecular defects observed in ciliopathies including Jeune syndrome, Ellis-van Creveld syndrome, short-rib polydactyly syndrome type 3, and orofaciodigital syndrome. The model enables examination of retrograde IFT deficiencies, protein accumulation at the ciliary tip, and disrupted GLI processing within a well-established epithelial framework.
Applications of the DYNC2LI1 Knockout HeLa Polyclonal Cells encompass ciliopathy disease modeling, hedgehog pathway dissection, cargo trafficking analysis, and drug screening for ciliary function restoration. Technical approaches include immunofluorescence with antibodies against ARL13B and acetylated tubulin to assess cilia, Western blotting for GLI proteins and dynein-2 components, RT-qPCR for hedgehog targets, and live-cell imaging of IFT dynamics. Co-immunoprecipitation can be used to investigate dynein-2 complex interactions. For further support, contact Ascent Research.
