Novel RNA Viral Vectors for Chemically Regulated Gene Expression in Embryonic Stem Cells
Novel RNA Viral Vectors for Chemically Regulated Gene Expression in Embryonic Stem Cells
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DOI：
10.1021/acssynbio.1c00214
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发表时间：
2021-11-19
影响因子：
4.7
通讯作者：
Yokobayashi, Yohei
中科院分区：
文献类型：
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作者：
Kim, Narae;Yokobayashi, Yohei
RNA病毒载体在没有DNA中间体的情况下复制，是生物医学和兽医应用中有吸引力的细胞操纵平台，因为它们具有最小的染色体整合风险。水泡性口炎病毒(VSV)载体是研究最多的RNA病毒载体之一，因为它们对人类的致病性较低，并且能够高水平表达转基因数周至数月。然而，由于它们的细胞致病性，它们的应用大多局限于溶瘤和疫苗载体。我们在VSV载体中发现了两个突变，它们协同增强了小鼠胚胎干细胞(ESCs)的稳定性，并显著降低了细胞病变效应。我们还展示了通过嵌入核糖开关对转基因表达的化学调节。感染突变载体的ESCs显示保持多能性。这一新载体为精确调控胚胎干细胞中的基因表达奠定了基础，从而在不改变染色体的情况下产生各种分化细胞。
RNA viral vectors that replicate without DNA intermediates are attractive platforms for manipulation of cells for biomedical and veterinary applications because they have minimal risk of chromosomal integration. Vesicular stomatitis virus (VSV) vectors are among the most well-studied RNA viral vectors due to their low pathogenicity to humans and ability to express transgenes at high levels for weeks to months. However, their applications have been mostly limited to oncolytic and vaccine vectors due to their cytopathogenicity. We discovered two mutations in the VSV vector that synergistically confer improved stability in mouse embryonic stem cells (ESCs) with markedly lower cytopathic effects. We also demonstrated chemical regulation of transgene expression through embedded riboswitches. The ESCs infected with the mutant vector were shown to maintain pluripotency. This new vector sets the stage for precise regulation of gene expression in ESCs to produce a variety of differentiated cells without chromosomal alteration.