Engineering RNA export for measurement and manipulation of living cells.

Engineering RNA export for measurement and manipulation of living cells.
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用于测量和操纵活细胞的工程RNA导出。

DOI:
10.1016/j.cell.2023.06.013
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发表时间:
2023-08-17
期刊:
影响因子:
64.5
通讯作者:
Elowitz, Michael B.
Elowitz, Michael B.
中科院分区:
生物学1区
文献类型:
--
作者:
Horns, Felix;Martinez, Joe A.;Fan, Chengcheng;Haque, Mehernaz;Linton, James M.;Tobin, Victoria;Santat, Leah;Maggiolo, Ailiena O.;Bjorkman, Pamela J.;Lois, Carlos;Elowitz, Michael B.

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从活细胞中可编程地输出RNA分子的系统将能够对细胞动力学进行非破坏性监测,并能够对能够向其他细胞提供可执行RNA程序的细胞进行工程。我们在病毒的启发下,开发出了基因编码的细胞RNA输出器,它能有效和选择性地将哺乳动物细胞中的靶RNA分子包装并分泌在保护性纳米颗粒中。通过导出和测序RNA条形码,可以通过克隆分辨率对细胞种群动态进行非破坏性监测。此外,通过在纳米颗粒中加入融合原,我们证明了输出的mRNA在受体细胞中的传递、表达和功能活性。我们称这些系统为Courier(用于询问、表达和调节的RNA的受控输出和摄取)。Courier能够测量细胞动力学,并为基于RNA细胞间传递的混合细胞和基因治疗奠定了基础。基于设计的蛋白质纳米笼,开发合成的胞外囊泡,用于可编程和高效的输出、细胞到细胞之间的输送和mRNA的表达。
A system for programmable export of RNA molecules from living cells would enable both non-destructive monitoring of cell dynamics and engineering of cells capable of delivering executable RNA programs to other cells. We developed genetically encoded cellular RNA exporters, inspired by viruses, that efficiently and selectively package and secrete target RNA molecules from mammalian cells within protective nanoparticles. Exporting and sequencing RNA barcodes enabled non-destructive monitoring of cell population dynamics with clonal resolution. Further, by incorporating fusogens into the nanoparticles, we demonstrated delivery, expression, and functional activity of exported mRNA in recipient cells. We term these systems COURIER (Controlled Output and Uptake of RNA for Interrogation, Expression, and Regulation). COURIER enables measurement of cell dynamics and establishes a foundation for hybrid cell and gene therapies based on cell-to-cell delivery of RNA. The development of synthetic extracellular vesicles, based on designed protein nanocages, for the programmable and efficient export, cell-to-cell delivery, and expression of mRNA.
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