RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells
文献类型：期刊论文
| 作者 | Basak E. Uygun; Yong Hu; Lei Li; Yin Yu; Haishui Huang; Martin L. Yarmush |
| 刊名 | Nano life
 |
| 出版日期 | 2018 |
| 文献子类 | 期刊论文 |
| 英文摘要 | Regenerative therapies require availability of an abundant healthy cell source which can be achieved by e±cient cryopreservation techniques. Here, we established a novel approach for improved cell cryopreservation using an mRNA-based dCas9-VP64 gene activation system for transient, yet highly e±cient expression of epigenetic related genes in mammalian cells for repression of metabolic activity. Before freezing, mammalian cells were treated by dCas9-VP64- modi¯ed mRNA and guide RNAs for upregulation of histone deacetylase (HDAC), DNA methyltransferase (DNMT) and transcriptional co-repressor Sin3A genes. Cell viability, karyotype, pluripotency, and other cell speci¯c functions were analyzed during post-thaw culture. Using conventional cryopreservation protocols, we found improvement of viability in dCas9- VP64 pretreated cells (P < 0:05) compared to untreated cells. Combined with dCas9-VP64 system, a reduced amount of cryoprotectant (5% DMSO) did not negatively a®ect the post-thaw viability. Co-elivering chemically modi¯ed dCas9-VP64 mRNA with gRNAs is an e±cient gene delivery method compared to DNA-based strategies, without the associated safety concerns. This approach is a simple, yet e®ective way to accelerate a wide array of cellular research and translational medical applications. |
| 语种 | 英语 |
| 源URL | [http://ir.siat.ac.cn:8080/handle/172644/13452]  |
| 专题 | 合成所 |
| 推荐引用方式 GB/T 7714 | Basak E. Uygun,Yong Hu,Lei Li,et al. RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells[J]. Nano life,2018. |
| APA | Basak E. Uygun,Yong Hu,Lei Li,Yin Yu,Haishui Huang,&Martin L. Yarmush.(2018).RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells.Nano life. |
| MLA | Basak E. Uygun,et al."RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells".Nano life (2018). |
入库方式： OAI收割
来源：深圳先进技术研究院
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