Potential Application of TALENs against Murine Cytomegalovirus Latent Infections
Potential Application of TALENs against Murine Cytomegalovirus Latent Infections
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DOI：
10.3390/v11050414
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发表时间：
2019-05-01
期刊：
影响因子：
4.7
通讯作者：
Chen, Yuan-Chuan
中科院分区：
文献类型：
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作者：
Chen, Shiu-Jau;Chen, Yuan-Chuan
巨细胞病毒（CMV）感染仍然是一个全球性的健康问题，因为潜伏病毒在人类中持续存在并引起复发性疾病。目前，没有针对巨细胞病毒潜伏感染的治疗方法，而针对活动性感染的治疗受到副作用和其他问题的限制。根除动物体内潜伏的病毒是不可能的。HCMV（人类巨细胞病毒）是人类疾病特有的；然而，由于HCMV的宿主特异性和较长的生命周期，研究难度较大。幸运的是，MCMV（小鼠巨细胞病毒）提供了一个很好的动物模型。本研究以MCMV M80/80.5序列为靶点，构建了三对特异性转录激活物样效应核酸酶（TALEN）质粒（MCMV1-2、3-4和5-6），以测试其在NIH3T3细胞培养中阻断MCMV裂解复制的效果。初步数据显示，在转染质粒之前，TALEN质粒对MCMV的M80/80.5序列具有特异性靶向和切割作用，能有效抑制MCMV在细胞培养中的生长。进一步利用最特异的TALEN质粒对（MCMV3-4）来证实其对Balb/c小鼠的潜在MCMV复制和基因表达的负调控作用。与对照组相比，注射特异性的TALEN质粒可显著抑制小鼠五个器官中立即早期基因（ie-1） DNA的拷贝数水平。结果表明，TALENs可能提供一种有效的策略来去除动物体内潜伏的MCMV。
Cytomegalovirus (CMV) infections are still a global health problem, because the latent viruses persist in humans and cause recurring diseases. Currently, there are no therapies for CMV latent infections and the therapies for active infections are limited by side effects and other problems. It is impossible to eradicate latent viruses in animals. HCMV (human CMV) is specific to human diseases; however, it is difficult to study HCMV due to its host specificity and long life cycle. Fortunately, MCMV (murine CMV) provides an excellent animal model. Here, three specific pairs of transcription activator-like effector nuclease (TALEN) plasmids (MCMV1-2, 3-4, and 5-6) were constructed to target the MCMV M80/80.5 sequence in order to test their efficacy in blocking MCMV lytic replication in NIH3T3 cell culture. The preliminary data showed that TALEN plasmids demonstrate specific targeting and cleavage in the MCMV M80/80.5 sequence and effectively inhibit MCMV growth in cell culture when the plasmid transfection is prior to the viral infection. The most specific pairs of TALEN plasmids (MCMV3-4) were further used to confirm the negative regulation of latent MCMV replication and gene expression in Balb/c mice. The injection of specific TALEN plasmids caused significant inhibition in the copy number level of immediately early gene (ie-1) DNA in five organs of mice, when compared with the controls. The result demonstrated that TALENs potentially provide an effective strategy to remove latent MCMV in animals.