CRISPR-Cas9 Screens Identify the RNA Helicase DDX3X as a Repressor of C9ORF72 (GGGGCC)n Repeat-Associated Non-AUG Translation

Cheng, Weiwei; Wang, Shaopeng; Zhang, Zhe; Morgens, David W.; Hayes, Lindsey R.; Lee, Soojin; Portz, Bede; Xie, Yuanyang; Nguyen, Baotram V.; Haney, Michael S.; Yan, Shirui; Dong, Daoyuan; Coyne, Alyssa N.; Yang, Junhua; Xian, Fengfan; Cleveland, Don W.; Qiu, Zhaozhu; Rothstein, Jeffrey D.; Shorter, James; Gao, Fen Biao; Bassik, Michael C.; Sun, Shuying

doi:10.1016/j.neuron.2019.09.003

Cited by 118 publications

(147 citation statements)

References 73 publications

Supporting

Mentioning

132

Contrasting

Order By: Relevance

“…Several of the identified genes have been previously linked to the pathogenesis of parkinsonisms and other neurodegenerative diseases. NFX1 has been identified through a CRISPR-Cas9 screen as a gene that reduces the levels of dipeptide repeat proteins (DPRs) by lowering the nuclear export of GGGGCCC repeat-containing transcripts upon silencing (Cheng et al, 2019;Hautbergue et al, 2017). That gene was also identified through our screen as a reducer of a-synuclein spread upon silencing, presumably through lowering intracellular levels of a-synuclein, thereby having a consistent directionality of effect as reported in that c9orf72 screen.…”

Section: Discussionsupporting

confidence: 63%

An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein

Kara

Crimi

Wiedmer

et al. 2019

Preprint

View full text Add to dashboard Cite

Neuropathological and experimental evidence suggests that the cell-to-cell transfer of a-synuclein has an important role in the pathogenesis of Parkinson's disease (PD). However, the mechanism underlying this phenomenon is not fully understood. We undertook an siRNA, genome-wide high throughput screen to identify genes regulating the cell-to-cell transfer of a-synuclein. We transiently transfected HEK cells stably overexpressing a-synuclein with a construct encoding GFP-2a-aSynuclein-RFP. The cells expressing a-synuclein-RFP through transfection were double positive for GFP and RFP fluorescence, whereas the cells receiving it through transfer were positive only for RFP fluorescence. The amount of asynuclein transfer was quantified by high content microscopy. A series of unbiased screens confirmed the involvement of 38 genes in the regulation of a-synuclein-RFP transfer. One of those hits was ITGA8, a candidate gene recently identified through a large PD genome wide association study (GWAS). Weighted gene co-expression network analysis (WGCNA) and weighted protein-protein network interaction analysis (WPPNIA) showed that the hits clustered in networks that included known PD Mendelian and GWAS risk genes more frequently than expected than random chance. Given the genetic overlap between a-synuclein transfer and PD, those findings provide supporting evidence for the importance of the cell-to-cell transfer of a-synuclein in the pathogenesis of PD, and expand our understanding of the mechanism of a-synuclein spread.

show abstract

Section: Discussionsupporting

confidence: 63%

An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein

Kara

Crimi

Wiedmer

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…5A-E). Recent studies have suggested that a reduction in DPR levels can mitigate stressor induced cell death and restore functional NCT in iPSNs (Cheng et al, 2019). However, this study did not evaluate the effects of DDX3X manipulation on NPC components themselves but was limited to the distribution of fluorescent shuttle proteins.…”

Section: Discussionmentioning

confidence: 99%

G₄C₂repeat RNA mediates the disassembly of the nuclear pore complex in C9orf72 ALS/FTD

Coyne

Zaepfel

Hayes

et al. 2020

Preprint

View full text Add to dashboard Cite

Nucleocytoplasmic transport, controlled by the nuclear pore complex, has recently emerged as a pathomechanism underlying neurodegenerative diseases including C9orf72 ALS/FTD. However, little is known about the underlying molecular events and the underlying biology in human neurons. Using super resolution structured illumination microscopy of twenty three nucleoporins in nuclei from C9orf72 iPSC derived neurons and postmortem human tissue we identify a unique subset of eight nucleoporins lost from human neuronal nuclei. POM121, an integral transmembrane nucleoporin, appears to coordinate the composition of the nucleoporins within human neuronal nuclei ultimately impacting nucleocytoplasmic transport, and subsequent cellular toxicity in C9orf72 iPSNs. These data suggest that POM121 is a critical nucleoporin in the maintenance of the nuclear localization of specific nucleoporins in human neurons. Moreover, loss of nuclear POM121, as a result of expanded C9orf72 ALS/FTD repeat RNA, initiates a pathological cascade affecting nucleoporin composition within neuronal nuclei, nuclear pore complex function, and overall downstream neuronal survival.

show abstract

“…Typically, potential therapies aim at the downstream effects arising from repeat expansion to reduce their toxic consequences. The goal is to either reverse gene silencing (70,81,469,(503)(504)(505) or alleviate adverse effects of toxic RNA (98, 101, 203, 506 -514) and/or toxic proteins (98,101,203,506,515). These methods, while worthy and promising, warrant several caveats.…”

Section: Can Reds Be Cured?mentioning

confidence: 99%

On the wrong DNA track: Molecular mechanisms of repeat-mediated genome instability

Khristich

Mirkin

2020

Journal of Biological Chemistry

232

239

View full text Add to dashboard Cite

Expansions of simple tandem repeats are responsible for almost 50 human diseases, the majority of which are severe, degenerative, and not currently treatable or preventable. In this review, we first describe the molecular mechanisms of repeat-induced toxicity, which is the connecting link between repeat expansions and pathology. We then survey alternative DNA structures that are formed by expandable repeats and review the evidence that formation of these structures is at the core of repeat instability. Next, we describe the consequences of the presence of long structure-forming repeats at the molecular level: somatic and intergenerational instability, fragility, and repeat-induced mutagenesis. We discuss the reasons for gender bias in intergenerational repeat instability and the tissue specificity of somatic repeat instability. We also review the known pathways in which DNA replication, transcription, DNA repair, and chromatin state interact and thereby promote repeat instability. We then discuss possible reasons for the persistence of disease-causing DNA repeats in the genome. We describe evidence suggesting that these repeats are a payoff for the advantages of having abundant simple-sequence repeats for eukaryotic genome function and evolvability. Finally, we discuss two unresolved fundamental questions: (i) why does repeat behavior differ between model systems and human pedigrees, and (ii) can we use current knowledge on repeat instability mechanisms to cure repeat expansion diseases?

show abstract

CRISPR-Cas9 Screens Identify the RNA Helicase DDX3X as a Repressor of C9ORF72 (GGGGCC)n Repeat-Associated Non-AUG Translation

Cited by 118 publications

References 73 publications

An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein

An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein

G₄C₂repeat RNA mediates the disassembly of the nuclear pore complex in C9orf72 ALS/FTD

On the wrong DNA track: Molecular mechanisms of repeat-mediated genome instability

Contact Info

Product

Resources

About

CRISPR-Cas9 Screens Identify the RNA Helicase DDX3X as a Repressor of C9ORF72 (GGGGCC)n Repeat-Associated Non-AUG Translation

Cited by 118 publications

References 73 publications

An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein

An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of a-synuclein

G4C2repeat RNA mediates the disassembly of the nuclear pore complex in C9orf72 ALS/FTD

On the wrong DNA track: Molecular mechanisms of repeat-mediated genome instability

Contact Info

Product

Resources

About

G₄C₂repeat RNA mediates the disassembly of the nuclear pore complex in C9orf72 ALS/FTD