Transgenic shRNA pigs reduce susceptibility to foot and mouth disease virus infection.
Transgenic shRNA pigs reduce susceptibility to foot and mouth disease virus infection.
复制标题
转基因 shRNA 猪可降低口蹄疫病毒感染的易感性。
DOI：
10.7554/elife.06951
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发表时间：
2015-06-19
期刊：
影响因子：
7.7
通讯作者：
Ouyang H
中科院分区：
文献类型：
--
作者：
Hu S;Qiao J;Fu Q;Chen C;Ni W;Wujiafu S;Ma S;Zhang H;Sheng J;Wang P;Wang D;Huang J;Cao L;Ouyang H
口蹄疫病毒（Foot-and-mouth disease virus，FMDV）是一种严重危害养猪业的病毒性疾病。一种新的替代策略是培育对感染具有遗传抗性的猪。在这里，我们生产的转基因（TG）猪组成型表达FMDV特异性短干扰RNA（siRNA）来自小发夹RNA（shRNA）。TG成纤维细胞的体外攻击显示shRNA抑制病毒生长。通过肌肉注射100 LD 50的FMDV攻击TG和非TG猪。在所有非TG猪中观察到高热、口蹄疫的严重临床体征和典型的组织病理学变化，但在高siRNA猪中没有观察到。我们的研究结果表明，TG shRNA可以提供一个可行的工具，生产动物与增强抗口蹄疫病毒。DOI：http://dx.doi.org/10.7554/eLife.06951.001口蹄疫经常在牲畜中引起严重的暴发。引起这种疾病的病毒可以感染牛、猪、绵羊、山羊和许多野生动物;这种疾病也具有高度传染性，传播非常迅速和容易。为了控制口蹄疫的传播，农民必须经常杀死已经暴露的整群动物。可能传播病毒的野生动物也可能被杀死，以阻止疾病的传播。预防口蹄疫的疫苗是可用的，通常用于帮助预防疾病的传播。然而，一旦口蹄疫开始爆发，疫苗阻止其传播可能为时已晚。这是因为疫苗可能需要大约一周的时间才能提供保护，而到那时，暴露的动物可能已经病得很重了。2010年进行的先前工作报告说，小鼠可以通过基因工程产生短链RNA分子，这些分子可以关闭口蹄疫病毒的基因。与感染了口蹄疫病毒的正常小鼠相比，基因工程小鼠体内几乎没有显示出这种疾病的迹象。现在，胡，乔，傅等人-包括一些参与2010年工作的研究人员，也以同样的方式对猪进行了基因改造。实验表明，当这些猪的细胞在实验室中暴露于口蹄疫病毒时，病毒的生长速度比正常情况下要慢得多。接下来，胡、乔、傅等人给转基因猪和非转基因猪注射病毒。所有正常的猪很快就出现了严重的症状，包括这种疾病特有的口腔和足部溃疡。此外，对这些猪的细胞的检查显示出疾病的迹象。但是基因工程猪并没有患上重病，它们的细胞几乎没有显示出疾病的迹象。一些转基因猪出现了一些溃疡，但这些溃疡出现的时间比正常情况要晚得多。到目前为止，研究结果表明，有可能培育出对口蹄疫有抵抗力的猪。胡、乔、傅等人下一步将确定转基因猪是否能将口蹄疫病毒传染给其他猪和牲畜。DOI：http://dx.doi.org/10.7554/eLife.06951.002网站
Foot-and-mouth disease virus (FMDV) is an economically devastating viral disease leading to a substantial loss to the swine industry worldwide. A novel alternative strategy is to develop pigs that are genetically resistant to infection. Here, we produce transgenic (TG) pigs that constitutively expressed FMDV-specific short interfering RNA (siRNA) derived from small hairpin RNA (shRNA). In vitro challenge of TG fibroblasts showed the shRNA suppressed viral growth. TG and non-TG pigs were challenged by intramuscular injection with 100 LD50 of FMDV. High fever, severe clinical signs of foot-and-mouth disease and typical histopathological changes were observed in all of the non-TG pigs but in none of the high-siRNA pigs. Our results show that TG shRNA can provide a viable tool for producing animals with enhanced resistance to FMDV. DOI: http://dx.doi.org/10.7554/eLife.06951.001 Foot-and-mouth disease regularly causes serious outbreaks in livestock. The virus that causes the disease can infect cattle, pigs, sheep, goats, and many species of wild animals; the disease is also highly contagious and spreads very quickly and easily. To control the spread of foot-and-mouth disease, farmers must often kill entire herds of animals that have been exposed. Wild animals that can spread the virus may also be killed in an effort to stop the spread of the disease. Vaccines that protect against foot-and-mouth disease are available and are often used to help prevent the spread of the disease. However, once an outbreak of foot-and-mouth disease begins it may be too late for vaccines to stop its spread. This is because the vaccines can take about a week to provide protection, and by that time an exposed animal may already be very ill. Previous work conducted in 2010 reported that mice could be genetically engineered to produce short stretches of RNA molecules that can switch off genes from the foot-and-mouth disease virus. Compared with normal mice infected with the foot-and-mouth disease virus, the genetically engineered mice showed little sign of the disease in their bodies. Now, Hu, Qiao, Fu et al.—including some of the researchers involved in the 2010 work—have genetically engineered pigs in the same way. The experiments show that when cells from these pigs are exposed to the foot-and-mouth disease virus in the laboratory, the virus grows much less than normal. Next, Hu, Qiao, Fu et al. injected genetically engineered pigs and non-genetically engineered pigs with the virus. All of the normal pigs developed severe symptoms very quickly, including the disease's characteristic mouth and foot sores. Additionally, examinations of these pigs' cells showed signs of the disease. But the genetically engineered pigs did not become seriously ill and their cells showed little sign of the disease. Some of the genetically engineered pigs developed a few sores but these sores appeared much later than normal. So far, the results suggest that it may be possible to develop pigs that are resistant to foot-and-mouth disease. Hu, Qiao, Fu et al. will next determine whether or not the genetically engineered pigs can pass the foot-and-mouth virus on to other pigs and livestock. DOI: http://dx.doi.org/10.7554/eLife.06951.002