Rosa26 creert2

Ptpn6^fl/fl, Cx3cr1-Cre, and Rosa26^LSL−YFP mice have been described elsewhere (25 (link), 35 (link), 36 (link)). Rosa26-CreERT2 and dLck-Cre mice were obtained from the Jackson Laboratory (37 (link), 38 (link)). C57BL/6 mice used for controls were obtained from Jackson Laboratories or Envigo. Mice were kept in specific pathogen-free facilities at the University of California, San Francisco (UCSF), Revolution Medicines or Taconic Biosciences, and cared for in accordance with institutional guidelines. All genotypes of mouse strains used were confirmed by PCR analysis of tail DNA.

Mice were bred and maintained at the Kansai Medical University Research Animal Facility in accordance with the Kansai Medical University guidelines. C57BL/6 J, Bmi1^CreER/+27 (link), Rosa26^CreERT2/+, and Rosa26^rbw/+ mice were purchased from Jackson Laboratories (Sacramento, CA, USA) or generated as previously described15 (link)16 (link)17 (link). The experiments were approved by the Kansai Medical University Welfare Committee. Tamoxifen (Sigma, St. Louis, MO, USA) was dissolved in corn oil (Sigma) and intraperitoneally injected into adult mice at concentrations of 9 and 5 mg/40 g body weight for Bmi1^CreER/+ mice and Rosa26^CreERT2/+ mice, respectively.

Recql4^K525A and Recql4^G522Efs mice were generated using Cripsr/Cas9 methods by the Mouse Engineering at Garvan/ABR (MEGA) services (Garvan Institute, Darlinghurst, Australia). Lysine 525 was mutated to Alanine (AAG>GCA) in single cell C57Bl/6 embryos via sgRNA-directed gene targeting and homologous recombination with a single stranded DNA oligonucleotide substrate. Viable pups were screened by DNA sequencing and one C57Bl/6 male carrying the K525A mutation on one allele and a 2bp insertion (GA) after the T521 codon (G522Efs) on the other allele was identified as a founder. The chemically (ENU) induced Recql4^M789K and Recql4^R347X mutations were obtained from the Australian Phenomics Facility (APF, Canberra, Australia: IGL01381 and IGL01809). Recql4^fl/fl mice (C57BL/6-Recql4^tm2272Arte) have been previously described [20 (link), 44 (link)]. Rosa26-CreER^T2 mice on a C57Bl/6 background were purchased from The Jackson Laboratory (B6.129-Gt(ROSA)26Sor^{tm1(cre/ERT2)Tyj}/J, Stock Number: 008463) and have been previously described [20 (link)]. All lines were on a backcrossed C57Bl/6 background. ENU mutants were outcrossed at least 6 times and assessed across multiple generations to eliminate effects of any additional mutations. Tamoxifen containing food was prepared by Specialty Feeds (Perth, Australia) at 400mg/kg Tamoxifen citrate (Sigma Aldrich) in a base of standard mouse chow.