RNA-sequencing of lung RNA from mice with compound mutations in ADAR1 and MAVS, ZBP1 or RIPK3.

The purpose of this study was to examine the role of MAVS, ZBP1 and RIPK3 in the phenotype that develops when ADAR1 activity is impaired, in particular when the Za domain of ADAR1 is mutated. Mice homozygous for a Za domain-mutant allele of Adar1 (Adar1mZa/mZa mice) and mice carrying one mZa and one null Adar1 allele (Adar1-/mZa mice) were compared with control mice that were either wild type or heterozygous for the Adar1 mZa allele (Adar1wt/mZa mice). The effects of MAVS deficiency, RIPK3 deficiency, ZBP1 deficiency or ZBP1 Za domain mutations were assessed by analysing compound mutant mice. Given the early postnatal lethal phenotype that develops in Adar1-/mZa mice, comparisons were made in RNA isolated from lung tissue from newborn mice of each genotype (5 mice per genotype). As Adar1-/mZa mice additionally lacking Mavs or Zbp1 are viable, adult mice (15-20 weeks of age) were also used for several compound mutations as donors of lung tissue.本研究旨在探究MAVS、ZBP1与RIPK3在ADAR1活性受损，尤其是ADAR1的Za结构域发生突变时所诱发的表型中的作用。本研究将携带Adar1的Za结构域突变等位基因的纯合小鼠（Adar1^mZa/mZa小鼠）、同时携带一个突变型Za等位基因与一个敲除型Adar1等位基因的小鼠（Adar1^-/mZa小鼠），与对照组小鼠（包括野生型或Adar1^mZa等位基因杂合小鼠，即Adar1^wt/mZa小鼠）进行对比。通过分析复合突变小鼠，本研究评估了MAVS缺陷、RIPK3缺陷、ZBP1缺陷或ZBP1的Za结构域突变所产生的生物学效应。鉴于Adar1^-/mZa小鼠会出现出生后早期致死的表型，研究人员对各基因型新生小鼠（每个基因型5只）的肺组织提取RNA开展了比较分析。由于同时缺失Mavs或Zbp1的Adar1^-/mZa小鼠可正常存活，研究人员还选取了15~20周龄的成年小鼠作为肺组织供体，用于部分复合突变相关实验。