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Wavefront Engineering Microscopy Group, Photonics Department, Institut de la Vision, Sorbonne Université, INSERM, CNRS, Paris, France.
Department of Biomedical Engineering, George Washington University, Washington, DC, USA.
Nat Rev Methods Primers. 2022;2. doi: 10.1038/s43586-022-00136-4. Epub 2022 Jul 21.
Optogenetic techniques have been developed to allow control over the activity of selected cells within a highly heterogeneous tissue, using a combination of genetic engineering and light. Optogenetics employs natural and engineered photoreceptors, mostly of microbial origin, to be genetically introduced into the cells of interest. As a result, cells that are naturally light-insensitive can be made photosensitive and addressable by illumination and precisely controllable in time and space. The selectivity of expression and subcellular targeting in the host is enabled by applying control elements such as promoters, enhancers and specific targeting sequences to the employed photoreceptor-encoding DNA. This powerful approach allows precise characterization and manipulation of cellular functions and has motivated the development of advanced optical methods for patterned photostimulation. Optogenetics has revolutionized neuroscience during the past 15 years and is primed to have a similar impact in other fields, including cardiology, cell biology and plant sciences. In this Primer, we describe the principles of optogenetics, review the most commonly used optogenetic tools, illumination approaches and scientific applications and discuss the possibilities and limitations associated with optogenetic manipulations across a wide variety of optical techniques, cells, circuits and organisms.
光遗传学技术已得到发展，可通过基因工程与光的结合，对高度异质组织内选定细胞的活动进行控制。光遗传学利用主要源自微生物的天然及工程化光感受器，通过基因手段导入目标细胞。这样一来，原本对光不敏感的细胞就能变得对光敏感，并可通过光照进行寻址，且在时间和空间上能得到精确控制。通过将启动子、增强子和特定靶向序列等控制元件应用于所使用的光感受器编码DNA，可实现宿主中表达的选择性和亚细胞靶向性。这种强大的方法能够精确表征和操纵细胞功能，并推动了用于图案化光刺激的先进光学方法的发展。在过去15年里，光遗传学彻底改变了神经科学，并且有望在包括心脏病学、细胞生物学和植物科学等其他领域产生类似的影响。在本入门文章中，我们描述光遗传学的原理，回顾最常用的光遗传学工具、光照方法和科学应用，并讨论在各种光学技术、细胞、回路和生物体中与光遗传学操作相关的可能性和局限性。
