Full-length cDNAs of Tmsb10 and Kras was amplified by PCR with sets of primers (Supplementary Table 7 ), and were used to construct p3xFLAG-TMSB10 and pCMV-HA-KRAS expression plasmids, respectively. The p3xFLAG-CMV10 expression plasmid (Sigma-Aldrich) was used as a control study.
To construct a donor plasmid for CRISPR/Cas9 technology, an 849-bp fragment upstream from the first ATG and an 849-bp fragment downstream from the first ATG of the Tmsb10 gene were amplified from the C57BL/6 genome. These fragments were used as the 5’ and 3’ homologous arms. mCherry tagged with human influenza hemagglutinin (HA) and Thoseaasigna virus 2 A (T2A) at the N- and C-terminal sites (HA-mCherry-T2A), respectively, was synthesized as follows. A DNA fragment encoding mCherry was amplified from pmCherry-N1 (Clontech, Palo Alto, CA, USA) and inserted into the EcoRI/BglII site of pCMV-HA (Takara, Shiga, Japan) to generate pCMV-HA-mCherry. Thereafter, the plasmid was subjected to PCR with a 3’ primer containing the T2A sequence to generate the HA-mCherry-T2A fragment. The primers used for amplification of the fragments are listed in Supplementary Table7 . Then, these three fragments (5’ homologous arm, HA-mCherry-T2A, and 3’ homologous arm) were inserted into the SalI/EcoRI site of the pBluescript II KS+ using an In-Fusion HD cloning kit (Clontech) (Supplementary Fig. 2 ).
To construct a donor plasmid for CRISPR/Cas9 technology, an 849-bp fragment upstream from the first ATG and an 849-bp fragment downstream from the first ATG of the Tmsb10 gene were amplified from the C57BL/6 genome. These fragments were used as the 5’ and 3’ homologous arms. mCherry tagged with human influenza hemagglutinin (HA) and Thoseaasigna virus 2 A (T2A) at the N- and C-terminal sites (HA-mCherry-T2A), respectively, was synthesized as follows. A DNA fragment encoding mCherry was amplified from pmCherry-N1 (Clontech, Palo Alto, CA, USA) and inserted into the EcoRI/BglII site of pCMV-HA (Takara, Shiga, Japan) to generate pCMV-HA-mCherry. Thereafter, the plasmid was subjected to PCR with a 3’ primer containing the T2A sequence to generate the HA-mCherry-T2A fragment. The primers used for amplification of the fragments are listed in Supplementary Table