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利用工程化黄热病疫苗 YFV-17D 进行顺行性跨神经元示踪和基因控制。

Anterograde transneuronal tracing and genetic control with engineered yellow fever vaccine YFV-17D.

机构信息

Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX, USA.

Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, TX, USA.

出版信息

Nat Methods. 2021 Dec;18(12):1542-1551. doi: 10.1038/s41592-021-01319-9. Epub 2021 Nov 25.

DOI:10.1038/s41592-021-01319-9
PMID:34824475
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8665090/
Abstract

Transneuronal viruses are powerful tools for tracing neuronal circuits or delivering genes to specific neurons in the brain. While there are multiple retrograde viruses, few anterograde viruses are available. Further, available anterograde viruses often have limitations such as retrograde transport, high neuronal toxicity or weak signals. We developed an anterograde viral system based on a live attenuated vaccine for yellow fever-YFV-17D. Replication- or packaging-deficient mutants of YFV-17D can be reconstituted in the brain, leading to efficient synapse-specific and anterograde-only transneuronal spreading, which can be controlled to achieve either monosynaptic or polysynaptic tracing. Moreover, inducible transient replication of YFV-17D mutant is sufficient to induce permanent transneuronal genetic modifications without causing neuronal toxicity. The engineered YFV-17D systems can be used to express fluorescent markers, sensors or effectors in downstream neurons, thus providing versatile tools for mapping and functionally controlling neuronal circuits.

摘要

神经病毒是追踪神经元回路或向大脑特定神经元传递基因的有力工具。虽然有多种逆行病毒，但可用的顺行病毒却很少。此外，现有的顺行病毒往往存在一些限制，如逆行运输、高神经元毒性或信号较弱。我们开发了一种基于减毒活疫苗黄热病毒（YFV-17D）的顺行病毒系统。YFV-17D 的复制或包装缺陷突变体能在大脑中重新构建，导致有效的突触特异性和仅顺行性的神经病毒传播，这种传播可以被控制，以实现单突触或多突触追踪。此外，YFV-17D 突变体的诱导性瞬时复制足以在不引起神经元毒性的情况下诱导永久性的神经病毒遗传修饰。工程化的 YFV-17D 系统可用于在下游神经元中表达荧光标记物、传感器或效应物，从而为映射和功能控制神经元回路提供了多功能工具。

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