Shcholok Tetiana, Eftekharpour Eftekhar
Spinal Cord Research Centre, Department of Physiology and Pathophysiology, University of Manitoba, Winnipeg, MB, Canada.
Neural Regen Res. 2023 Feb;18(2):273-279. doi: 10.4103/1673-5374.346541.
Gene deletion has been a valuable tool for unraveling the mysteries of molecular biology. Early approaches included gene trapping and gene targetting to disrupt or delete a gene randomly or at a specific location, respectively. Using these technologies in mouse embryos led to the generation of mouse knockout models and many scientific discoveries. The efficacy and specificity of these approaches have significantly increased with the advent of new technology such as clustered regularly interspaced short palindromic repeats for targetted gene deletion. However, several limitations including unwanted off-target gene deletion have hindered their widespread use in the field. Cre-recombinase technology has provided additional capacity for cell-specific gene deletion. In this review, we provide a summary of currently available literature on the application of this system for targetted deletion of neuronal genes. This article has been constructed to provide some background information for the new trainees on the mechanism and to provide necessary information for the design, and application of the Cre-recombinase system through reviewing the most frequent promoters that are currently available for genetic manipulation of neurons. We additionally will provide a summary of the latest technological developments that can be used for targeting neurons. This may also serve as a general guide for the selection of appropriate models for biomedical research.
基因缺失一直是解开分子生物学奥秘的重要工具。早期方法包括基因捕获和基因靶向，分别用于随机或在特定位置破坏或删除基因。在小鼠胚胎中使用这些技术催生了小鼠基因敲除模型，并带来了许多科学发现。随着成簇规律间隔短回文重复序列等新技术的出现，用于靶向基因缺失，这些方法的有效性和特异性显著提高。然而，包括意外的脱靶基因缺失在内的一些局限性阻碍了它们在该领域的广泛应用。Cre重组酶技术为细胞特异性基因缺失提供了更多能力。在这篇综述中，我们总结了目前关于该系统用于神经元基因靶向缺失应用的现有文献。本文旨在为新学员提供该机制的一些背景信息，并通过回顾目前可用于神经元基因操作的最常用启动子，为Cre重组酶系统的设计和应用提供必要信息。我们还将总结可用于靶向神经元的最新技术发展。这也可作为生物医学研究中选择合适模型的一般指南。
