Mitochondrial supercomplexes are functionally important, but how supercomplexes being assembled is not known. Mitochondrial Complex I has an important role in the assembly of supercomplex. In this study, we generated a series of cell model with shRNA mediated gene silencing or CRISPR-Cas9 gene knockout technology. We performed a series of mitochondrial proteome analyses in different cell lines with disruption of gene that encodes subunit of Complex I. Also, we performed Co-IP of exogenous TIMMDC1, which was an assembly factor of Complex I.