The CRISPR/Cas9-derived Ift140 knockout cells were no longer able to form cilia (Fig. 3b).
ExperimentName: Mouse ciliated cell model of Ift140 loss-of-function
CellLine: Mouse Inner Medullary Collecting Duct 3 (mIMCD3) cells
CellEFO: EFO:0002887 (mouse cell line), EFO:0001643 (kidney-derived cell line)
GeneAlteration: The cell line was subjected to CRISPR-based knockout of endogenous IFT140.
CellPhenotype: Ift140 knockout cells exhibited a complete inability to form cilia relative to the control rescued by overexpression of wild-type Ift140 (Figure 3b).
CellEvidence: The defect in ciliary formation induced by Ift140 knockout (Figure 3b) recapitulates one of the cellular features of human patients harboring IFT140 variants (absence of cilia from a high proportion of fibroblasts). This also matches the Chlamydomonas model of IFT140 loss described in PMID: 28207750, albeit with greater severity.
CellVariant: CRISPR-based knockout was performed in this study to mimic IFT140 loss of function, while the human IFT140 variants were tested in the related rescue experiment instead.