The nedd-8 activating enzyme gene underlies genetic resistance to infectious pancreatic necrosis virus in Atlantic salmon.
The nedd-8 activating enzyme gene underlies genetic resistance to infectious pancreatic necrosis virus in Atlantic salmon.
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DOI：
10.1016/j.ygeno.2021.09.012
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发表时间：
2021-11
期刊：
影响因子：
4.4
通讯作者：
Houston RD
中科院分区：
文献类型：
--
作者：
Pavelin J;Jin YH;Gratacap RL;Taggart JB;Hamilton A;Verner-Jeffreys DW;Paley RK;Rubin CJ;Bishop SC;Bron JE;Robledo D;Houston RD
大西洋鲑鱼对传染性胰腺坏死病毒（IPNV）的遗传抗性是一个罕见的例子，其中一个单一的基因座（QTL）解释了几乎所有的遗传变异。基于鲑鱼26号染色体上的这个QTL的遗传标记测试已被广泛应用于选择性育种，以显着降低疾病的发病率。在本研究中，全基因组测序和功能注释的方法被应用到QTL区域中的p53基因和变体。这是通过分析鲑鱼鱼苗的纯合抗性和纯合易感基因型与IPNV挑战之间的差异表达的补充。这些分析指出，NEDD-8激活酶1（nae 1）基因作为一个假定的功能候选QTL效应的基础。通过CRISPR-Cas9敲除大西洋鲑鱼细胞系中的nae 1基因和化学抑制nae 1蛋白活性，进一步评估了nae 1在IPN抗性中的作用，这两者都导致生产性IPNV复制的高度显著减少。相比之下，CRISPR-Cas9敲除先前被认为是病毒细胞受体的候选基因（cdh 1）对IPNV的生产性复制没有重大影响。这些结果表明，nae 1是影响鲑鱼IPNV抗性的主要QTL的致病基因，为neddylation在宿主-病原体相互作用中的关键作用提供了进一步的证据，并强调了将高通量基因组学方法与靶向基因组编辑相结合以了解抗病性的遗传基础的价值。利用全基因组测序和功能注释技术对大西洋鲑IPNV抗性QTL进行定位。基因表达分析指出，NEDD-8激活酶1（nae 1）作为一个假定的功能候选人的QTL。CRISPR-Cas9敲除和细胞系中nae 1的化学抑制导致IPNV复制显著减少。CRISPR-Cas9敲除先前提出的候选基因（cdh 1）对IPNV的生产性复制没有影响。这些结果表明，nae 1是致病基因的主要QTL影响鲑鱼IPNV的抗性。
Genetic resistance to infectious pancreatic necrosis virus (IPNV) in Atlantic salmon is a rare example of a trait where a single locus (QTL) explains almost all of the genetic variation. Genetic marker tests based on this QTL on salmon chromosome 26 have been widely applied in selective breeding to markedly reduce the incidence of the disease. In the current study, whole genome sequencing and functional annotation approaches were applied to characterise genes and variants in the QTL region. This was complemented by an analysis of differential expression between salmon fry of homozygous resistant and homozygous susceptible genotypes challenged with IPNV. These analyses pointed to the NEDD-8 activating enzyme 1 (nae1) gene as a putative functional candidate underlying the QTL effect. The role of nae1 in IPN resistance was further assessed via CRISPR-Cas9 knockout of the nae1 gene and chemical inhibition of the nae1 protein activity in Atlantic salmon cell lines, both of which resulted in highly significant reduction in productive IPNV replication. In contrast, CRISPR-Cas9 knockout of a candidate gene previously purported to be a cellular receptor for the virus (cdh1) did not have a major impact on productive IPNV replication. These results suggest that nae1 is the causative gene underlying the major QTL affecting resistance to IPNV in salmon, provide further evidence for the critical role of neddylation in host-pathogen interactions, and highlight the value in combining high-throughput genomics approaches with targeted genome editing to understand the genetic basis of disease resistance. Whole genome sequencing and functional annotation were used to characterise the major IPNV resistance QTL in Atlantic salmon. Gene expression analyses pointed to NEDD-8 activating enzyme 1 (nae1) as a putative functional candidate underlying the QTL. CRISPR-Cas9 knockout and chemical inhibition of nae1 in cell lines resulted in significant reduction in IPNV replication. CRISPR-Cas9 knockout of a previously suggested candidate gene (cdh1) did not have an impact on productive IPNV replication. These results suggest that nae1 is the causative gene underlying the major QTL affecting resistance to IPNV in salmon.
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影响因子：
1.2
作者：
Cingolani, Pablo;Platts, Adrian;Ruden, Douglas M.
通讯作者：
Ruden, Douglas M.
影响因子：
16
作者：
Jin YH;Robledo D;Hickey JM;McGrew MJ;Houston RD
通讯作者：
Houston RD
DOI：
10.1007/s10126-020-09995-y
发表时间：
2020-10
期刊：
Marine biotechnology (New York, N.Y.)
影响因子：
--
作者：
Gratacap RL;Jin YH;Mantsopoulou M;Houston RD
通讯作者：
Houston RD
影响因子：
4.4
作者：
Moen T;Baranski M;Sonesson AK;Kjøglum S
通讯作者：
Kjøglum S
影响因子：
7
作者：
Rise, ML;von Schalburg, KR;Koop, BF
通讯作者：
Koop, BF