B6.129x1 sarm1tm1aidi j

All experiments were conducted with the approval of the Johns Hopkins Animal Care and Use Committee. C3-PMP mice (B6.Cg-Tg(PMP22)C3Fbas/J, referred to as CMT1A mice) were obtained from the Jackson Laboratory, expanded by mating CMT1A females with wildtype males (C57BL/6J) and genotyped according to the suggested protocol. SARM1 knockout mice (B6.129X1-Sarm1^tm1Aidi/J) were also obtained from the Jackson Laboratory and genotyped according to the suggested protocol. CMT1A/SARM1^+/− mice were generated by first mating CMT1A mice with SARM1 knockout mice and then mating CMT1A/SARM1^+/− mice with wildtype/SARM1 ^+/− mice. A balanced representation of male and female mice aged approximately three and six months were assessed by SHIRPA and inverted holding test three times per day for three consecutive days (days 1–3) followed by evaluation with an accelerating rotarod assay and grip strength test three times per day for three consecutive days (days 4–6) and nerve electrophysiology and tissue harvest was performed on day 7. The Hargreaves test was performed on a separate batch of three-month-old mice.

Mice carrying a null allele of Sarm1 (B6.129X1-Sarm1^tm1Aidi/J) were obtained from the Jackson Laboratories (Stock Number: 018069). The Wallerian Degeneration Slow (Wld^S) allele (Lunn et al., 1989 (link); Mack et al., 2001 ) was backcrossed into C57BL/6J > 20 generations. DR6^−/− mice (Tnfrsf21^tm2Gne) were obtained from Genentech (Zhao et al., 2001 (link)). To genetically label RGC axons for histological assessment of morphological signs of axon degeneration, Thy1-CFP mice (B6.Cg-Tg(Thy1-CFP)23Jrs/J, Stock Number: 003710) which express CFP in ~80% of RGCs(Bernstein et al., 2007 (link)) were crossed to Sarm1^−/− and Wld^S mice. Sarm1^−/− and Wld^S mice were crossed to generate Sarm1^−/−Wld^S double mutants. SpCas9 knockin mice (Rosa26 locus) were obtained from Jackson labs (stock 026179). Either littermate controls that did not carry any deleted allele or the Wld^S allele or C57BL6/J mice were used as experimental wildtype (WT) controls. Mice were housed in a 12-hour light dark cycle and were fed chow and water ad libitum. All experiments were conducted in accordance with the Association for Research in Vision and Ophthalmology’s statement on the use of animals in ophthalmic research and were approved by the University of Rochester’s Committee on Animal Resources and the Johns Hopkins University School of Medicine Institutional Animal Care and Use Committee.