当前位置: X-MOL 学术Cell Rep. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Conditional Single Vector CRISPR/SaCas9 Viruses for Efficient Mutagenesis in the Adult Mouse Nervous System.
Cell Reports ( IF 7.5 ) Pub Date : 2020-03-24 , DOI: 10.1016/j.celrep.2020.02.092
Avery C Hunker 1 , Marta E Soden 2 , Dasha Krayushkina 3 , Gabriel Heymann 3 , Rajeshwar Awatramani 4 , Larry S Zweifel 1
Affiliation  

Mice engineered for conditional, cell type-specific gene inactivation have dominated the field of mouse genetics because of the high efficiency of Cre-loxP-mediated recombination. Recent advances in CRISPR/Cas9 technologies have provided alternatives for rapid gene mutagenesis for loss-of-function (LOF) analysis. Whether these strategies can be streamlined for rapid genetic analysis with the efficiencies comparable with those of conventional genetic approaches has yet to be established. We show that a single adeno-associated viral (AAV) vector containing a recombinase-dependent Staphylococcus aureus Cas9 (SaCas9) and a single guide RNA (sgRNA) are as efficient as conventional conditional gene knockout and can be adapted for use in either Cre- or Flp-driver mouse lines. The efficacy of this approach is demonstrated for the analysis of GABAergic, glutamatergic, and monoaminergic neurotransmission. Using this strategy, we reveal insight into the role of GABAergic regulation of midbrain GABA-producing neurons in psychomotor activation.

中文翻译:

在成年小鼠神经系统中有效诱变的条件单载体CRISPR / SaCas9病毒。

由于Cre-loxP介导的重组效率高,为条件性,细胞类型特异性基因失活而设计的小鼠主导了小鼠遗传学领域。CRISPR / Cas9技术的最新进展为功能丧失(LOF)分析的快速基因诱变提供了替代方法。是否可以简化这些策略以进行快速的遗传分析,其效率可与传统遗传方法相媲美。我们显示,包含重组酶依赖性金黄色葡萄球菌Cas9(SaCas9)和单个指导RNA(sgRNA)的单个腺相关病毒(AAV)载体与常规条件基因敲除一样有效,并且可以适用于Cre-或Flp驱动程序鼠标线。这种方法的功效已证明可用于分析GABA能,谷氨酸能和单胺能神经传递。使用这种策略,我们揭示了对中脑产生GABA的神经元的GABA能调节在精神运动激活中的作用的见解。
更新日期:2020-03-26
down
wechat
bug