These DYNC2LI1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the DYNC2LI1 gene in HEK293T cells. This polyclonal population offers a versatile loss-of-function model to study the functional role of the DYNC2LI1-encoded light intermediate chain subunit of the cytoplasmic dynein-2 complex, which is critical for retrograde intraflagellar transport (IFT) and primary cilium-dependent signaling.
HEK293T cells are immortalized human embryonic kidney epithelial cells transformed with SV40 large T antigen, renowned for their high transfection efficiency and robust protein expression capabilities. Widely utilized for viral packaging and heterologous expression, HEK293T cells can also be induced to form primary cilia, rendering them a practical and genetically tractable model for ciliary biology and intracellular trafficking studies.
DYNC2LI1 functions as a light intermediate chain of the cytoplasmic dynein-2 motor complex, which is essential for retrograde IFT??the process by which ciliary tip-derived signaling components and IFT machinery are transported back to the cell body. The gene is transcriptionally regulated by ciliogenic factors such as RFX3 and FOXJ1 and acts downstream of Sonic hedgehog (SHH) ligand stimulation. DYNC2LI1-mediated transport is required for proper processing of GLI2 and GLI3 transcription factors; disruption leads to attenuated expression of Hedgehog target genes including PTCH1, GLI1, and HHIP. The encoded protein directly interacts with dynein-2 heavy chain DYNC2H1, intermediate chain WDR34, light chains DYNLRB1 and DYNLL1, and the dynein-2 accessory factor WDR60, as well as with IFT complex A and B components along the microtubules of the axoneme.
In the HEK293T background, knockout of DYNC2LI1 impairs retrograde ciliary transport without affecting global cell viability, permitting direct assessment of cilia-dependent signaling defects. Because this polyclonal population retains endogenous expression of core Hedgehog pathway components??including PTCH1, SMO, SUFU, PKA, CK1, and GSK3??it is particularly well-suited for interrogating the biochemical consequences of disrupted dynein-2 function. The model??s amenability to high-efficiency transfection enables complementation with wild-type or mutant DYNC2LI1 constructs, facilitating structure?Cfunction analyses. Notably, biallelic loss-of-function mutations in DYNC2LI1 cause the human ciliopathy short-rib thoracic dysplasia 15 (SRTD15), also known as Jeune syndrome, making this cellular system relevant for understanding pathogenic mechanisms driving skeletal ciliopathies.
Researchers can deploy these polyclonal knockout cells in a variety of experimental workflows: ciliogenesis assays combined with immunofluorescence to quantify cilium length and morphology; co-immunoprecipitation and mass spectrometry to map the dynein-2 interactome; GLI luciferase reporter assays to measure Hedgehog pathway activity; RT-qPCR or RNA-seq to profile transcriptomic changes; and drug sensitivity screens aimed at identifying compounds that restore ciliary trafficking or rescue downstream signaling. The polyclonal nature allows robust population-level measurements while avoiding clonal selection artifacts. For further technical specifications or to inquire about custom gene-editing services, please contact Ascent Research.
