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Infectivity assessment of porcine endogenous retrovirus using high-throughput sequencing technologies
Biologicals ( IF 1.5 ) Pub Date : 2021-05-24 , DOI: 10.1016/j.biologicals.2021.05.001
Ken Kono 1 , Kiyoko Kataoka 1 , Yuzhe Yuan 2 , Keisuke Yusa 2 , Kazuhisa Uchida 2 , Yoji Sato 3
Affiliation  

Xenogenic cell-based therapeutic products are expected to alleviate the chronic shortage of human donor organs. For example, porcine islet cell products are currently under development for the treatment of human diabetes. As porcine cells possess endogenous retrovirus (PERV), which can replicate in human cells in vitro, the potential transmission of PERV has raised concerns in the case of products that use living pig cells as raw materials. Although several PERV sequences exist in the porcine genome, not all have the ability to infect human cells. Therefore, polymerase chain reaction analysis, which amplifies a portion of the target gene, may not accurately assess the infection risk. Here, we determined porcine genome sequences and evaluated the infectivity of PERVs using high-throughput sequencing technologies. RNA sequencing was performed on both PERV-infected human cells and porcine cells, and reads mapped to PERV sequences were examined. The normalized number of the reads mapped to PERV regions was able to predict the infectivity of PERVs, indicating that it would be useful for evaluation of the PERV infection risk prior to transplantation of porcine products.



中文翻译:

使用高通量测序技术评估猪内源性逆转录病毒的感染性

预计基于异种细胞的治疗产品将缓解人类供体器官的长期短缺。例如,目前正在开发用于治疗人类糖尿病的猪胰岛细胞产品。由于猪细胞具有内源性逆转录病毒(PERV),它可以在体外在人类细胞复制,在使用活猪细胞作为原材料的产品的情况下,PERV 的潜在传播引起了人们的关注。尽管猪基因组中存在多个 PERV 序列,但并非所有序列都具有感染人类细胞的能力。因此,扩增部分目标基因的聚合酶链反应分析可能无法准确评估感染风险。在这里,我们确定了猪基因组序列并使用高通量测序技术评估了 PERV 的感染性。对受 PERV 感染的人类细胞和猪细胞进行 RNA 测序,并检查映射到 PERV 序列的读数。映射到 PERV 区域的标准化读数数量能够预测 PERV 的感染性,

更新日期:2021-06-05
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