Generation and Propagation of Genetically Engineered Mice

Kras^LSL-G12D/+ mice^{30 (link)} were a gift from T. Jacks (Massachusetts Institute of Technology). Rosa26^{LSL-Cas9-EGFP/+} mice^{31 (link)} were obtained from Jackson labs. Rosa26^{LoxP-Cas9-EGFP/+} mice were generated by methods described previously^{15 (link)}. Athymic nude (nu/nu) mice (5 to 6 weeks old) for the allograft study were purchased from Taconic Biosciences and maintained in Duke University’s accredited animal facility. MEFs were transduced with lentivirus and 2 million cells were injected into the hind limbs of nude mice. All animal studies were performed in accordance with protocols approved by the Duke University Institutional Animal Care and Use Committee (IACUC).