Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation.
Using the CRISPR/Cas9 system to understand neuropeptide biology and regulation.
复制标题
DOI：
10.1016/j.npep.2016.11.010
复制
发表时间：
2017-08
期刊：
影响因子：
2.9
通讯作者：
MacKenzie A
中科院分区：
文献类型：
--
作者：
Hay EA;Knowles C;Kolb A;MacKenzie A
神经肽及其受体在食欲、应激和炎症性疼痛等生理反应中发挥作用。由于神经肽具有如此多样化和重要的生理作用，敲除编码它们的基因、它们的受体、它们的部分调控序列，或复制与疾病相关的多态变异是研究神经肽及其如何导致疾病的重要步骤。以前，基因敲除是通过胚胎干细胞的靶向同源重组等方法产生的，但这种方法既昂贵又耗时。CRISPR/Cas9系统已经迅速占领了基因组编辑领域，并将促进我们对神经肽基因及其调控的理解。利用CRISPR/Cas9技术，可以大大减少制作转基因动物模型所需的时间和成本。在这篇综述中，我们描述了该系统如何通过细胞系或动物模型中的“敲除”或“敲入”突变来操纵基因组序列。我们还讨论了系统的特殊性和限制脱靶效应的方法。当结合基因组序列的可用性时，CRISPR/Cas9指导的体外和体内基因组编辑有望比以往任何时候都更深入地了解健康和疾病中神经肽的生物学。CRISPR/Cas9系统旨在促进我们对神经肽基因的理解。CRISPR/Cas9系统允许高效的基因组编辑。敲除突变比敲入突变更有效。CRISPR/Cas9的脱靶效应可以通过不同的策略进行控制。
Neuropeptides and their receptors play a role in physiological responses such as appetite, stress and inflammatory pain. With neuropeptides having such diverse and important physiological roles, knocking-out the genes encoding them, their receptors, parts of their regulatory sequences, or reproducing disease associated polymorphic variants are important steps in studying neuropeptides and how they may contribute to disease. Previously, knock-outs were generated using methods such as targeted homologous recombination in embryonic stem cells but this method is costly and time-consuming. The CRISPR/Cas9 system has rapidly taken over the genome editing field and will advance our understanding of neuropeptide genes and their regulation. With CRISPR/Cas9 technology, the time and costs involved in producing transgenic animal models, is greatly reduced. In this review, we describe how the system can be used to manipulate genomic sequences by “knock-out” or “knock-in” mutations in cell lines or in animal models. We also discuss the specificity of the system and methods to limit off-target effects. When combined with the availability of genome sequences, CRISPR/Cas9 directed genome editing in vitro and in vivo, promises to provide a deeper understanding of the biology of the neuropeptides in health and disease than has ever been available before. The CRISPR/Cas9 system is set to advance our understanding of neuropeptide genes. The CRISPR/Cas9 system allows for efficient genome editing. Knock-out mutations can be produced more efficiently than knock-in mutations. Off-target effects of CRISPR/Cas9 can be controlled using different strategies.
登录
查看更多内容
影响因子：
4.6
作者：
Inui M;Miyado M;Igarashi M;Tamano M;Kubo A;Yamashita S;Asahara H;Fukami M;Takada S
通讯作者：
Takada S
影响因子：
4.6
作者：
Horii T;Arai Y;Yamazaki M;Morita S;Kimura M;Itoh M;Abe Y;Hatada I
通讯作者：
Hatada I
影响因子：
48
作者：
Brown, Andrew J.;Fisher, Daniel A.;Cui, Xiaoxia
通讯作者：
Cui, Xiaoxia
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle