Scaled and Efficient Derivation of Loss of Function Alleles in Risk Genes for Neurodevelopmental and Psychiatric Disorders in Human iPSC.
| العنوان: | Scaled and Efficient Derivation of Loss of Function Alleles in Risk Genes for Neurodevelopmental and Psychiatric Disorders in Human iPSC. |
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| المؤلفون: | Zhang H; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., Peyton L; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., McCarroll A; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., de León Guerrerro SD; Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ.; Child Health Institute of New Jersey, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ., Zhang S; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL.; Department of Psychiatry and Behavioral Neuroscience, The University of Chicago, Chicago, IL., Gowda P; Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ.; Child Health Institute of New Jersey, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ., Sirkin D; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., El Achwah M; Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ.; Child Health Institute of New Jersey, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ., Duhe A; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., Wood WG; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., Jamison B; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., Tracy G; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL., Pollak R; Center for Advanced Biotechnology and Medicine, Rutgers Robert Wood Johnson Medical School, Piscataway, NJ., Hart RP; Department of Cell Biology and Neuroscience, Rutgers University., Pato CN; Center for Advanced Biotechnology and Medicine, Rutgers Robert Wood Johnson Medical School, Piscataway, NJ., Mulle JG; Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ.; Center for Advanced Biotechnology and Medicine, Rutgers Robert Wood Johnson Medical School, Piscataway, NJ.; Department of Psychiatry, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ., Sanders AR; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL.; Department of Psychiatry and Behavioral Neuroscience, The University of Chicago, Chicago, IL., Pang ZP; Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ.; Child Health Institute of New Jersey, Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ., Duan J; Center for Psychiatric Genetics, NorthShore University HealthSystem, Evanston, IL.; Department of Psychiatry and Behavioral Neuroscience, The University of Chicago, Chicago, IL. |
| المصدر: | BioRxiv : the preprint server for biology [bioRxiv] 2024 Mar 19. Date of Electronic Publication: 2024 Mar 19. |
| نوع المنشور: | Preprint |
| اللغة: | English |
| بيانات الدورية: | Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: | Translating genetic findings for neurodevelopmental and psychiatric disorders (NPD) into actionable disease biology would benefit from large-scale and unbiased functional studies of NPD genes. Leveraging the cytosine base editing (CBE) system, here we developed a pipeline for clonal loss-of-function (LoF) allele mutagenesis in human induced pluripotent stem cells (hiPSCs) by introducing premature stop-codons (iSTOP) that lead to mRNA nonsense-mediated-decay (NMD) or protein truncation. We tested the pipeline for 23 NPD genes on 3 hiPSC lines and achieved highly reproducible, efficient iSTOP editing in 22 NPD genes. Using RNAseq, we confirmed their pluripotency, absence of chromosomal abnormalities, and NMD. Interestingly, for three schizophrenia risk genes ( SETD1A, TRIO , CUL1 ), despite the high efficiency of base editing, we only obtained heterozygous LoF alleles, suggesting their essential roles for cell growth. We replicated the reported neural phenotypes of SHANK3 -haploinsufficiency and found CUL1 -LoF reduced neurite branches and synaptic puncta density. This iSTOP pipeline enables a scaled and efficient LoF mutagenesis of NPD genes, yielding an invaluable shareable resource. Competing Interests: Declaration of Interests The authors declare no conflict of interests. |
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| معلومات مُعتمدة: | R01 MH116281 United States MH NIMH NIH HHS; R01 MH106575 United States MH NIMH NIH HHS; R01 MH125528 United States MH NIMH NIH HHS; R01 AA023797 United States AA NIAAA NIH HHS; R01 AG063175 United States AG NIA NIH HHS; RM1 MH133065 United States MH NIMH NIH HHS |
| تواريخ الأحداث: | Date Created: 20240402 Latest Revision: 20240408 |
| رمز التحديث: | 20240408 |
| مُعرف محوري في PubMed: | PMC10983959 |
| DOI: | 10.1101/2024.03.18.585542 |
| PMID: | 38562852 |
| قاعدة البيانات: | MEDLINE |
| DOI: | 10.1101/2024.03.18.585542 |
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