*4.2. RNA Interference and CRISPR/Cas9 System*

For RAB10 silencing experiments, anti-RAB10 siRNA (ON TARGETplus human RAB10 siRNA SMARTpool) and scramble control siRNA (ON TARGETplus non-targeting control pool) from Horizon Discovery (Cambridge, UK) were used. RAB10-knockout (KO) HEK cells were established using the CRISPR/Cas9 technology as previously described [46]. Briefly, sgRNA guide sequences targeting human RAB10 were designed using the CRISPR design tool from Horizon Discovery (sense, 5′–GCGTACGTCTTCTTCGCCATT–3′ and antisense, 5′–AATGGCGAAGAAGACGTACGC–3′ ) and cloned into pSpCas9(BB)-2A-Puro plasmid (PX459, Addgene). After sequence verification, the resulting plasmid was transfected into HEK cells, and 24 h later, 2 µg/mL puromycin (Gibco) was added to the culture medium for the selection of transfected cells.
