Next-Generation Sequencing to compare RNA transcripts between Adam19 wildtype and knockout mice

ADAM19 has been associated with pulmonary function traits in a number of genome wide association analyses by our group and others. We want to use a mouse model to examine the role Adam19 in lung function to clarify whether ADAM19 is truly the causal gene at this locus. We created a whole body Adam19 knockout mouse model targeting exon6 and 7 to disrupt the transcript expression from exon 8 through the end of gene sequences. In previous publications, mice lacking Adam19 exon 11, which harbor a catalytic site, were not postnatal viable. Contradictory to the previous publications, our Adam19 knockout survived postnatally. Our objectives were to determine if this novel mouse model is truly Adam19-deficient rather than a hypomorph or neomorph rescuing the previously published lethal phenotype of Adam19-deficient mice. mRNA profiles of the heart and lung of wildtype and Adam19 knockout generated by deep sequencing, in triplicate, using Illumina NextSeq-High.本团队及其他研究者已在多项全基因组关联分析（Genome-Wide Association Analysis, GWAS）中，将ADAM19与肺功能表型建立了关联。本研究拟通过小鼠模型探究Adam19在肺功能调控中的作用，以明确ADAM19是否为该基因座的真正致病基因。我们通过靶向外显子6（exon6）与外显子7（exon7），构建了全身性Adam19基因敲除小鼠模型，以破坏从外显子8（exon8）至基因序列末端的转录本表达。既往研究显示，缺失携带催化结构域（catalytic site）的Adam19外显子11（exon11）的小鼠无法在出生后存活，但本研究构建的Adam19基因敲除小鼠可在出生后正常存活，与既往研究结果相悖。本研究的核心目标为验证该新型小鼠模型是否为真正的Adam19缺陷型模型，而非通过低功能突变体（hypomorph）或新功能突变体（neomorph）挽救了既往报道的Adam19缺陷小鼠致死表型。本研究采用Illumina NextSeq-High平台，对野生型（wildtype）及Adam19基因敲除小鼠的心脏、肺组织进行深度测序（deep sequencing），每组设置三次生物学重复，以获取mRNA表达谱数据。