Removal of a partial genomic duplication restores synaptic transmission and behavior in the MyosinVA mutant mouse Flailer.
Removal of a partial genomic duplication restores synaptic transmission and behavior in the MyosinVA mutant mouse Flailer.
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DOI：
10.1186/s12915-023-01714-y
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发表时间：
2023-11-14
期刊：
影响因子：
5.4
通讯作者：
Constantine-Paton, Martha
中科院分区：
文献类型：
--
作者：
Bustos, Fernando J;Pandian, Swarna;Haensgen, Henny;Zhao, Jian-Ping;Strouf, Haley;Heidenreich, Matthias;Swiech, Lukasz;Deverman, Benjamin E;Gradinaru, Viviana;Zhang, Feng;Constantine-Paton, Martha
拷贝数变异，特别是基因组区域的重复，与包括自闭症谱系障碍(ASD)在内的各种神经退行性疾病密切相关。这些基因变异已被发现对大脑发育和功能有重大影响，可能会导致神经和行为症状的出现。开发针对这些基因组复制的策略一直是具有挑战性的，因为重复基因的内源拷贝的存在通常会使编辑策略复杂化。使用ASD和焦虑小鼠模型Flailer，其中包含部分基因组复制作为MyoVa的显性阴性，我们演示了在体外和体内使用DN-CRISPR来移除700hp的基因组区域。重要的是，dN-CRISPR还没有被用来使用偏移超过300bp的sgRNA来移除基因组区域。我们发现，编辑初级皮质神经元中的Flailer基因可以逆转突触运输和传输缺陷。此外，在羽翼动物身上中断的长期抑郁(LTD)在基因编辑后得到恢复。体内递送DN-CRISPR表明，局部递送到腹侧海马区可以挽救一些突变的行为，而脑室内递送可以完全恢复与焦虑和ASD相关的闪动动物的表型。我们的研究结果表明，DN-CRISPR有可能有效地去除较大的基因组重复，成为治疗神经退行性疾病的一种新的基因治疗方法。网上版载有补充材料，可在10.1186/s12915-023-01714-y查阅。
Copy number variations, and particularly duplications of genomic regions, have been strongly associated with various neurodegenerative conditions including autism spectrum disorder (ASD). These genetic variations have been found to have a significant impact on brain development and function, which can lead to the emergence of neurological and behavioral symptoms. Developing strategies to target these genomic duplications has been challenging, as the presence of endogenous copies of the duplicate genes often complicates the editing strategies. Using the ASD and anxiety mouse model Flailer, which contains a partial genomic duplication working as a dominant negative for MyoVa, we demonstrate the use of DN-CRISPRs to remove a 700 bp genomic region in vitro and in vivo. Importantly, DN-CRISPRs have not been used to remove genomic regions using sgRNA with an offset greater than 300 bp. We found that editing the flailer gene in primary cortical neurons reverts synaptic transport and transmission defects. Moreover, long-term depression (LTD), disrupted in Flailer animals, is recovered after gene editing. Delivery of DN-CRISPRs in vivo shows that local delivery to the ventral hippocampus can rescue some of the mutant behaviors, while intracerebroventricular delivery, completely recovers the Flailer animal phenotype associated to anxiety and ASD. Our results demonstrate the potential of DN-CRISPR to efficiently remove larger genomic duplications, working as a new gene therapy approach for treating neurodegenerative diseases. The online version contains supplementary material available at 10.1186/s12915-023-01714-y.