Genome-wide synthetic lethal CRISPR screen identifies FIS1 as a genetic interactor of ALS-linked C9ORF72
Open accessPublished version of a preprint
Abstract
Mutations in the C9ORF72 gene are the most common cause of amyotrophic lateral sclerosis (ALS). Both toxic gain of function and loss of function pathogenic mechanisms have been proposed. Accruing evidence from mouse knockout studies point to a role for C9ORF72 as a regulator of immune function. To provide further insight into its cellular function, we performed a genome-wide synthetic lethal CRISPR screen in human myeloid cells lacking C9ORF72. We discovered a strong synthetic lethal genetic interaction between C9ORF72 and FIS1, which encodes a mitochondrial membrane protein involved in mitochondrial fission and mitophagy. Mass spectrometry experiments revealed that in C9ORF72 knockout cells, FIS1 strongly bound to a class of immune regulators that activate the receptor for advanced glycation end (RAGE) products and trigger inflammatory cascades. These findings present a novel genetic interactor for C9ORF72 and suggest a compensatory role for FIS1 in suppressing inflammatory signaling in the absence of C9ORF72.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
iPSC-derived cells from different lineages can be combined into defined three-dimensional cultures called assembloids.
“Furthermore, iPSC-derived cell types from different lineages can be combined into defined three-dimensional cultures, termed assembloids9,94–98.”
From CRISPR-based functional genomics for neurological disease (2020) · cites this paper as [94]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2
Love, M. I., Huber, W., & Anders, S. (2014) · Genome Biology · doi:10.1186/s13059-014-0550-8 · Cited by 95,106 · see citing works
Zhang, Y., Chen, K., Sloan, S. A., Bennett, M. L., Scholze, A. R., O'Keeffe, S., Phatnani, H. P., Guarnieri, P., Caneda, C., Ruderisch, N., et al. (2014) · The Journal of Neuroscience · doi:10.1523/jneurosci.1860-14.2014 · Cited by 5,508 · see citing works
Genome-scale CRISPR-Cas9 knockout screening in human cells
Shalem, O., Sanjana, N. E., Hartenian, E., Shi, X., Scott, D. A., Mikkelsen, T. S., Heckl, D., Ebert, B. L., Root, D. E., Doench, J. G., et al. (2014) · Science · doi:10.1126/science.1247005 · Cited by 5,234 · see citing works
DeJesus-Hernandez, M., Mackenzie, I. R., Boeve, B. F., Boxer, A. L., Baker, M., Rutherford, N. J., Nicholson, A. M., Finch, N. A., Flynn, H., Adamson, J., et al. (2011) · Neuron · doi:10.1016/j.neuron.2011.09.011 · Cited by 4,936 · see citing works
A Hexanucleotide Repeat Expansion in C9ORF72 Is the Cause of Chromosome 9p21-Linked ALS-FTD
Renton, A. E., Majounie, E., Waite, A., Simón-Sánchez, J., Rollinson, S., Gibbs, J. R., Schymick, J. C., Laaksovirta, H., van Swieten, J. C., Myllykangas, L., et al. (2011) · Neuron · doi:10.1016/j.neuron.2011.09.010 · Cited by 4,409 · see citing works
Genetic screens in human cells using the CRISPR-Cas9 system
Wang, T., Wei, J. J., Sabatini, D. M., & Lander, E. S. (2014) · Science · doi:10.1126/science.1246981 · Cited by 2,856 · see citing works
Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation
Gilbert, L. A., Horlbeck, M. A., Adamson, B., Villalta, J. E., Chen, Y., Whitehead, E. H., Guimaraes, C., Panning, B., Ploegh, H. L., Bassik, M. C., et al. (2014) · Cell · doi:10.1016/j.cell.2014.09.029 · Cited by 2,728 · see citing works
Self-citationLi, W., Xu, H., Xiao, T., Cong, L., Love, M. I., Zhang, F., Irizarry, R. A., Liu, J. S., Brown, M., & Liu, X. S. (2014) · Genome Biology · doi:10.1186/s13059-014-0554-4 · Cited by 2,684 · see citing works
Easy quantitative assessment of genome editing by sequence trace decomposition
Brinkman, E. K., Chen, T., Amendola, M., & van Steensel, B. (2014) · Nucleic Acids Research · doi:10.1093/nar/gku936 · Cited by 2,390 · see citing works
Zhang, Y., Sloan, S. A., Clarke, L. E., Caneda, C., Plaza, C. A., Blumenthal, P. D., Vogel, H., Steinberg, G. K., Edwards, M. S. B., Li, G., et al. (2016) · Neuron · doi:10.1016/j.neuron.2015.11.013 · Cited by 2,295 · see citing works
Systematic Genetic Analysis with Ordered Arrays of Yeast Deletion Mutants
Tong, A. H. Y., Evangelista, M., Parsons, A. B., Xu, H., Bader, G. D., Pagé, N., Robinson, M., Raghibizadeh, S., Hogue, C. W. V., Bussey, H., et al. (2001) · Science · doi:10.1126/science.1065810 · Cited by 2,138 · see citing works
CHOPCHOP v3: expanding the CRISPR web toolbox beyond genome editing
Labun, K., Montague, T. G., Krause, M., Cleuren, Y. N. T., Tjeldnes, H., & Valen, E. (2019) · Nucleic Acids Research · doi:10.1093/nar/gkz365 · Cited by 2,009 · see citing works
The CRAPome: a contaminant repository for affinity purification–mass spectrometry data
Mellacheruvu, D., Wright, Z., Couzens, A. L., Lambert, J.-P., St-Denis, N. A., Li, T., Miteva, Y. V., Hauri, S., Sardiu, M. E., Low, T. Y., et al. (2013) · Nature Methods · doi:10.1038/nmeth.2557 · Cited by 1,686 · see citing works
Wang, S., Song, R., Wang, Z., Jing, Z., Wang, S., & Ma, J. (2018) · Frontiers in Immunology · doi:10.3389/fimmu.2018.01298 · Cited by 1,450 · see citing works
Koike-Yusa, H., Li, Y., Tan, E.-P., Velasco-Herrera, M. D. C., & Yusa, K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2800 · Cited by 1,064 · see citing works
Mitochondria–lysosome contacts regulate mitochondrial fission via RAB7 GTP hydrolysis
Wong, Y. C., Ysselstein, D., & Krainc, D. (2018) · Nature · doi:10.1038/nature25486 · Cited by 885 · see citing works
Foell, D., Wittkowski, H., Vogl, T., & Roth, J. (2006) · Journal of Leukocyte Biology · doi:10.1189/jlb.0306170 · Cited by 877 · see citing works
Ryckman, C., Vandal, K., Rouleau, P., Talbot, M., & Tessier, P. A. (2003) · The Journal of Immunology · doi:10.4049/jimmunol.170.6.3233 · Cited by 861 · see citing works
Yoon, Y., Krueger, E. W., Oswald, B. J., & McNiven, M. A. (2003) · Molecular and Cellular Biology · doi:10.1128/mcb.23.15.5409-5420.2003 · Cited by 820 · see citing works
Principles for the buffering of genetic variation
Hartman, J. L., Garvik, B., & Hartwell, L. (2001) · Science · doi:10.1126/science.1056072 · Cited by 808 · see citing works
Principles for the Buffering of Genetic Variation
Hartman IV, J. L. (2001) · Science · doi:10.1126/science.291.5506.1001 · Cited by 808 · see citing works
High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells
Zhou, Y., Zhu, S., Cai, C., Yuan, P., Li, C., Huang, Y., & Wei, W. (2014) · Nature · doi:10.1038/nature13166 · Cited by 733 · see citing works
Mozdy, A. D., McCaffery, J. M., & Shaw, J. M. (2000) · The Journal of Cell Biology · doi:10.1083/jcb.151.2.367 · Cited by 725 · see citing works
hFis1, a Novel Component of the Mammalian Mitochondrial Fission Machinery
James, D. I., Parone, P. A., Mattenberger, Y., & Martinou, J. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m303758200 · Cited by 653 · see citing works
Haploinsufficiency leads to neurodegeneration in C9ORF72 ALS/FTD human induced motor neurons
Shi, Y., Lin, S., Staats, K. A., Li, Y., Chang, W.-H., Hung, S.-T., Hendricks, E., Linares, G. R., Wang, Y., Son, E. Y., et al. (2018) · Nature Medicine · doi:10.1038/nm.4490 · Cited by 566 · see citing works
<i>C9orf72</i> is required for proper macrophage and microglial function in mice
O’Rourke, J. G., Bogdanik, L., Yáñez, A., Lall, D., Wolf, A. J., Muhammad, A. K. M. G., Ho, R., Carmona, S., Vit, J. P., Zarrow, J., et al. (2016) · Science · doi:10.1126/science.aaf1064 · Cited by 550 · see citing works
Fis1 and Bap31 bridge the mitochondria–ER interface to establish a platform for apoptosis induction
Iwasawa, R., Mahul‐Mellier, A., Datler, C., Pazarentzos, E., & Grimm, S. (2011) · The EMBO Journal · doi:10.1038/emboj.2010.346 · Cited by 520 · see citing works
Binding of S100 proteins to RAGE: An update
Leclerc, E., Fritz, G., Vetter, S. W., & Heizmann, C. W. (2008) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/j.bbamcr.2008.11.016 · Cited by 490 · see citing works
S100A8/A9: From basic science to clinical application
Pruenster, M., Vogl, T., Roth, J., & Sperandio, M. (2016) · Pharmacology & Therapeutics · doi:10.1016/j.pharmthera.2016.07.015 · Cited by 451 · see citing works
RAGE regulation and signaling in inflammation and beyond
Kierdorf, K., & Fritz, G. (2013) · Journal of Leukocyte Biology · doi:10.1189/jlb.1012519 · Cited by 434 · see citing works
The C9orf72 protein interacts with Rab1a and the ULK1 complex to regulate initiation of autophagy
Webster, C. P., Smith, E. F., Bauer, C. S., Moller, A., Hautbergue, G. M., Ferraiuolo, L., Myszczynska, M. A., Higginbottom, A., Walsh, M. J., Whitworth, A. J., et al. (2016) · The EMBO Journal · doi:10.15252/embj.201694401 · Cited by 404 · see citing works
Systematic comparison of CRISPR/Cas9 and RNAi screens for essential genes
Morgens, D. W., Deans, R. M., Li, A., & Bassik, M. C. (2016) · Nature Biotechnology · doi:10.1038/nbt.3567 · Cited by 384 · see citing works
Self-citationGenome-scale measurement of off-target activity using Cas9 toxicity in high-throughput screens
Morgens, D. W., Wainberg, M., Boyle, E. A., Ursu, O., Araya, C. L., Tsui, C. K., Haney, M. S., Hess, G. T., Han, K., Jeng, E. E., et al. (2017) · Nature Communications · doi:10.1038/ncomms15178 · Cited by 345 · see citing works
Self-citationLevels of human Fis1 at the mitochondrial outer membrane regulate mitochondrial morphology
Stojanovski, D., Koutsopoulos, O. S., Okamoto, K., & Ryan, M. T. (2004) · Journal of Cell Science · doi:10.1242/jcs.01058 · Cited by 332 · see citing works
Sunahori, K., Yamamura, M., Yamana, J., Takasugi, K., Kawashima, M., Yamamoto, H., Chazin, W., Nakatani, Y., Yui, S., & Makino, H. (2006) · Arthritis Research & Therapy · doi:10.1186/ar1939 · Cited by 324 · see citing works
Levine, T. P., Daniels, R. D., Gatta, A. T., Wong, L. H., & Hayes, M. J. (2013) · Bioinformatics · doi:10.1093/bioinformatics/bts725 · Cited by 321 · see citing works
Synthetic lethality: General principles, utility and detection using genetic screens in human cells
Nijman, S. M. B. (2010) · FEBS Letters · doi:10.1016/j.febslet.2010.11.024 · Cited by 313 · see citing works
Mitochondrial Rab GAPs govern autophagosome biogenesis during mitophagy
Yamano, K., Fogel, A. I., Wang, C., van der Bliek, A. M., & Youle, R. J. (2014) · eLife · doi:10.7554/elife.01612 · Cited by 307 · see citing works
Sullivan, P. M., Zhou, X., Robins, A. M., Paushter, D. H., Kim, D., Smolka, M. B., & Hu, F. (2016) · Acta Neuropathologica Communications · doi:10.1186/s40478-016-0324-5 · Cited by 295 · see citing works
Waite, A. J., Bäumer, D., East, S. Z., Neal, J., Morris, H. R., Ansorge, O., & Blake, D. J. (2014) · Neurobiology of Aging · doi:10.1016/j.neurobiolaging.2014.01.016 · Cited by 277 · see citing works
Loss-of-function mutations in the <i>C9ORF72</i> mouse ortholog cause fatal autoimmune disease
Burberry, A., Suzuki, N., Wang, J.-Y., Moccia, R., Mordes, D. A., Stewart, M. H., Suzuki-Uematsu, S., Ghosh, S., Singh, A., Merkle, F. T., et al. (2016) · Science Translational Medicine · doi:10.1126/scitranslmed.aaf6038 · Cited by 269 · see citing works
Atanasio, A., Decman, V., White, D., Ramos, M., Ikiz, B., Lee, H.-C., Siao, C.-J., Brydges, S., LaRosa, E., Bai, Y., et al. (2016) · Scientific Reports · doi:10.1038/srep23204 · Cited by 263 · see citing works
Human Fis1 regulates mitochondrial dynamics through inhibition of the fusion machinery
Yu, R., Jin, S., Lendahl, U., Nistér, M., & Zhao, J. (2019) · The EMBO Journal · doi:10.15252/embj.201899748 · Cited by 251 · see citing works
A C9ORF72/SMCR8-containing complex regulates ULK1 and plays a dual role in autophagy
Yang, M., Liang, C., Swaminathan, K., Herrlinger, S., Lai, F., Shiekhattar, R., & Chen, J.-F. (2016) · Science Advances · doi:10.1126/sciadv.1601167 · Cited by 238 · see citing works
Kramer, N. J., Haney, M. S., Morgens, D. W., Jovičić, A., Couthouis, J., Li, A., Ousey, J., Ma, R., Bieri, G., Tsui, C. K., et al. (2018) · Nature Genetics · doi:10.1038/s41588-018-0070-7 · Cited by 226 · see citing works
Self-citationMutations in Fis1 disrupt orderly disposal of defective mitochondria
Shen, Q., Yamano, K., Head, B. P., Kawajiri, S., Cheung, J. T. M., Wang, C., Cho, J.-H., Hattori, N., Youle, R. J., & van der Bliek, A. M. (2014) · Molecular Biology of the Cell · doi:10.1091/mbc.e13-09-0525 · Cited by 209 · see citing works
Calcium-dependent Tetramer Formation of S100A8 and S100A9 is Essential for Biological Activity
Leukert, N., Vogl, T., Strupat, K., Reichelt, R., Sorg, C., & Roth, J. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.04.009 · Cited by 195 · see citing works
Identification of phagocytosis regulators using magnetic genome-wide CRISPR screens
Haney, M. S., Bohlen, C. J., Morgens, D. W., Ousey, J. A., Barkal, A. A., Tsui, C. K., Ego, B. K., Levin, R., Kamber, R. A., Collins, H., et al. (2018) · Nature Genetics · doi:10.1038/s41588-018-0254-1 · Cited by 193 · see citing works
Self-citationC9orf72 binds SMCR8, localizes to lysosomes, and regulates mTORC1 signaling
Amick, J., Roczniak-Ferguson, A., & Ferguson, S. M. (2016) · Molecular Biology of the Cell · doi:10.1091/mbc.e16-01-0003 · Cited by 189 · see citing works
Loss of C9orf72 Enhances Autophagic Activity via Deregulated mTOR and TFEB Signaling
Ugolino, J., Ji, Y. J., Conchina, K., Chu, J., Nirujogi, R. S., Pandey, A., Brady, N. R., Hamacher-Brady, A., & Wang, J. (2016) · PLOS Genetics · doi:10.1371/journal.pgen.1006443 · Cited by 185 · see citing works
A global analysis of genetic interactions in Caenorhabditis elegans
Byrne, A. B., Weirauch, M. T., Wong, V., Koeva, M., Dixon, S. J., Stuart, J. M., & Roy, P. J. (2007) · Journal of Biology · doi:10.1186/jbiol58 · Cited by 172 · see citing works
Onoue, K., Jofuku, A., Ban-Ishihara, R., Ishihara, T., Maeda, M., Koshiba, T., Itoh, T., Fukuda, M., Otera, H., Oka, T., et al. (2012) · Journal of Cell Science · doi:10.1242/jcs.111211 · Cited by 160 · see citing works
There has been an awakening: Emerging mechanisms of C9orf72 mutations in FTD/ALS
Gitler, A. D., & Tsuiji, H. (2016) · Brain Research · doi:10.1016/j.brainres.2016.04.004 · Cited by 152 · see citing works
Self-citationC9ORF72 interaction with cofilin modulates actin dynamics in motor neurons
Sivadasan, R., Hornburg, D., Drepper, C., Frank, N., Jablonka, S., Hansel, A., Lojewski, X., Sterneckert, J. L., Hermann, A., Shaw, P. J., et al. (2016) · Nature Neuroscience · doi:10.1038/nn.4407 · Cited by 148 · see citing works
Sudria-Lopez, E., Koppers, M., de Wit, M., van der Meer, C., Westeneng, H.-J., Zundel, C. A. C., Youssef, S. A., Harkema, L., de Bruin, A., Veldink, J. H., et al. (2016) · Acta Neuropathologica · doi:10.1007/s00401-016-1581-x · Cited by 130 · see citing works
Rouhiainen, A., Kuja‐Panula, J., Tumova, S., & Rauvala, H. (2012) · Methods in molecular biology · doi:10.1007/978-1-62703-230-8_15 · Cited by 112 · see citing works
Maharjan, N., Künzli, C., Buthey, K., & Saxena, S. (2016) · Molecular Neurobiology · doi:10.1007/s12035-016-9850-1 · Cited by 99 · see citing works
Insights into structural and regulatory roles of Sec16 in COPII vesicle formation at ER exit sites
Yorimitsu, T., & Sato, K. (2012) · Molecular Biology of the Cell · doi:10.1091/mbc.e12-05-0356 · Cited by 89 · see citing works
Jorch, S. K., Austermann, J., Papantonopoulou, O., Riemenschneider, M., Xue, J., Bertheloot, D., Freise, N., Spiekermann, C., Witten, A., Viemann, D., et al. (2014) · The Journal of Immunology · doi:10.4049/jimmunol.1401085 · Cited by 78 · see citing works
Jofuku, A., Ishihara, N., & Mihara, K. (2005) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2005.05.154 · Cited by 73 · see citing works
ULK1 phosphorylates Sec23A and mediates autophagy-induced inhibition of ER-to-Golgi traffic
Gan, W., Zhang, C., Siu, K. Y., Satoh, A., Tanner, J. A., & Yu, S. S. B. (2017) · BMC Cell Biology · doi:10.1186/s12860-017-0138-8 · Cited by 52 · see citing works
Włodkowic, D., Faley, S. L., Darżynkiewicz, Z., et al. (2011) · Methods in molecular biology · doi:10.1007/978-1-61779-080-5_23 · Cited by 28 · see citing works
Edgar, K. A., Belvin, M. P., Parks, A. L., Whittaker, K. L., Mahoney, M. B., Nicoll, M., Park, C. C., Winter, C. G., Chen, F., Lickteig, K., et al. (2005) · Genetics · doi:10.1534/genetics.104.036343 · Cited by 18 · see citing works
Time-Lapse Imaging of Cell DeathPreprint
Wållberg, F., Tenev, T., & Meier, P. (2016) · Cold Spring Harbor Protocols · doi:10.1101/pdb.prot087395 · Cited by 18 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
CRISPR-based functional genomics for neurological disease
Kampmann, M. (2020) · Nature Reviews Neurology · doi:10.1038/s41582-020-0373-z · Cited by 163 · see citing works
Loss of <i>Fis1</i> impairs proteostasis during skeletal muscle aging in <i>Drosophila</i>
Lee, T., Chen, P., Su, M. P., Li, J., Chang, Y., Liu, R., Juan, H., Yang, J., Chan, S., Tsai, Y., et al. (2021) · Aging Cell · doi:10.1111/acel.13379 · Cited by 27 · see citing works
- Authors
- Chai, N., Haney, M. S., Couthouis, J., Morgens, D. W., Benjamin, A., Wu, K., Ousey, J., Fang, S., Finer, S., Bassik, M. C., & Gitler, A. D.
- Affiliations
- Stanford Medicine (US)
- Corresponding author
- Gitler, Aaron D.
- ORCID
- Venue
- Brain Research
- Publisher
- Elsevier BV
- Published
- 2020-02
- Volume
- 1728
- Pages
- 146601
- ISSN
- 0006-8993 (ISSN-L)
- PMC
- PMC7539795
- OpenAlex
- W2995406300
- Cited by
- 26
- References
- 64
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 3 · 2023: 4 · 2022: 3 · 2021: 8 · 2020: 4
- FWCI
- 2.54
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- Brain Research (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Neurological Disorders and Stroke
- Topics
- Health Sciences › Medicine › Neurology › Amyotrophic Lateral Sclerosis Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Keywords
- C9orf72, amyotrophic lateral sclerosis, Fis1, synthetic lethality, CRISPR screen, mitochondrial fission, mitophagy, RAGE, advanced glycation end products, inflammation, immune regulation, myeloid cells
- MeSH terms
- Synthetic Lethal Mutations, C9orf72 Protein, RNA-Seq, Amyotrophic Lateral Sclerosis, Genetic Testing, Humans, Membrane Proteins, U937 Cells, Mitochondrial Proteins, Gene Knockout Techniques, Clustered Regularly Interspaced Short Palindromic Repeats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
