For this end, we injected adeno-associated virus (AAV) encoding wild-type TMEM25 or two TMEM25 mutants (R326W and L338F), which were found in human colorectal cancers but not in human breast cancers in the catalogue of somatic mutations in cancer (COSMIC) database (https://cancer.sanger.ac.uk/cosmic), into mammary fat pad of 8-week-old female MMTV-PyMT transgenic mice. Here, TMEM25 is linked to breast cancer.