CRISPR interference to evaluate modifiers of C9ORF72-mediated toxicity in FTD.
CRISPR interference to evaluate modifiers of C9ORF72-mediated toxicity in FTD.
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CRISPR干扰以评价FTD中C9ORF72介导毒性的修饰剂。
DOI：
10.3389/fcell.2023.1251551
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发表时间：
2023
影响因子：
5.5
通讯作者：
Petrucelli, Leonard
中科院分区：
文献类型：
--
作者：
Pickles, Sarah;Alepuz, Desiree Zanetti;Koike, Yuka;Yue, Mei;Tong, Jimei;Liu, Pinghu;Zhou, Yugui;Jansen-West, Karen;Daughrity, Lillian M. M.;Song, Yuping;DeTure, Michael;Oskarsson, Bjoern;Graff-Radford, Neill R. R.;Boeve, Bradley F. F.;Petersen, Ronald C. C.;Josephs, Keith A. A.;Dickson, Dennis W. W.;Ward, Michael E. E.;Dong, Lijin;Prudencio, Mercedes;Cook, Casey N. N.;Petrucelli, Leonard
神经退行性疾病的治疗，包括额颞叶痴呆（FTD）和肌萎缩侧索硬化症（ALS），仍然相当有限，强调需要更大的机制洞察力和疾病相关模型。我们开发遗传风险因子、疾病修饰因子和其他FTD/ALS相关靶点的新型疾病模型的能力受到开发常规敲除和转基因小鼠所需的大量时间和资金的阻碍。为了克服这些限制，我们已经产生了一种新的CRISPRi干扰（CRISPRi）敲入小鼠。CRISPRi使用Cas9的催化死亡形式，融合到转录阻遏物以敲低蛋白质表达，然后引入针对感兴趣基因的单向导RNA。为了验证该模型的实用性，我们选择了TAR DNA结合蛋白（TDP-43）剪接靶，stathmin-2（STMN 2）。由于TDP-43活性丧失，FTD/ALS中STMN 2 RNA下调，STMN 2丧失在ALS发病机制中发挥作用。STMN 2功能丧失与FTD的关系尚未确定。我们发现，家族性FTD病例中的STMN 2蛋白水平与对照组相比显著降低，支持STMN 2缺失可能参与FTD的发病机制。在这里，我们提供了概念验证，我们可以同时敲低Stmn 2和表达9号染色体开放阅读框72（C9 ORF 72）基因中的扩展重复序列，成功复制C9相关病理学特征。有趣的是，Stmn 2的缺失对二肽重复蛋白（DPRs）的表达或沉积没有影响，但显著降低了磷酸化Tdp-43（pTdp-43）内含物的数量。我们提出，我们的新型CRISPRi小鼠提供了一种通用和快速的方法来沉默体内基因表达，并提出这种模型将有助于了解孤立或其他神经退行性疾病模型背景下的基因功能。
Treatments for neurodegenerative disease, including Frontotemporal dementia (FTD) and Amyotrophic lateral sclerosis (ALS), remain rather limited, underscoring the need for greater mechanistic insight and disease-relevant models. Our ability to develop novel disease models of genetic risk factors, disease modifiers, and other FTD/ALS-relevant targets is impeded by the significant amount of time and capital required to develop conventional knockout and transgenic mice. To overcome these limitations, we have generated a novel CRISPRi interference (CRISPRi) knockin mouse. CRISPRi uses a catalytically dead form of Cas9, fused to a transcriptional repressor to knockdown protein expression, following the introduction of single guide RNA against the gene of interest. To validate the utility of this model we have selected the TAR DNA binding protein (TDP-43) splicing target, stathmin-2 (STMN2). STMN2 RNA is downregulated in FTD/ALS due to loss of TDP-43 activity and STMN2 loss is suggested to play a role in ALS pathogenesis. The involvement of STMN2 loss of function in FTD has yet to be determined. We find that STMN2 protein levels in familial FTD cases are significantly reduced compared to controls, supporting that STMN2 depletion may be involved in the pathogenesis of FTD. Here, we provide proof-of-concept that we can simultaneously knock down Stmn2 and express the expanded repeat in the Chromosome 9 open reading frame 72 (C9ORF72) gene, successfully replicating features of C9-associated pathology. Of interest, depletion of Stmn2 had no effect on expression or deposition of dipeptide repeat proteins (DPRs), but significantly decreased the number of phosphorylated Tdp-43 (pTdp-43) inclusions. We submit that our novel CRISPRi mouse provides a versatile and rapid method to silence gene expression in vivo and propose this model will be useful to understand gene function in isolation or in the context of other neurodegenerative disease models.
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影响因子：
4
作者：
Kampmann M
通讯作者：
Kampmann M
影响因子：
64.5
作者：
CHUNG, JH;WHITELEY, M;FELSENFELD, G
通讯作者：
FELSENFELD, G
影响因子：
16.2
作者：
Cheng, Weiwei;Wang, Shaopeng;Sun, Shuying
通讯作者：
Sun, Shuying
影响因子：
15.1
作者：
Chew, Jeannie;Cook, Casey;Petrucelli, Leonard
通讯作者：
Petrucelli, Leonard
DOI：
10.1126/science.aaa9344
发表时间：
2015-06-05
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chew J;Gendron TF;Prudencio M;Sasaguri H;Zhang YJ;Castanedes-Casey M;Lee CW;Jansen-West K;Kurti A;Murray ME;Bieniek KF;Bauer PO;Whitelaw EC;Rousseau L;Stankowski JN;Stetler C;Daughrity LM;Perkerson EA;Desaro P;Johnston A;Overstreet K;Edbauer D;Rademakers R;Boylan KB;Dickson DW;Fryer JD;Petrucelli L
通讯作者：
Petrucelli L