Parallel in vivo analysis of large-effect autism genes implicates cortical neurogenesis and estrogen in risk and resilience

Neuron, Год журнала: 2021, Номер 109(5), С. 788 - 804.e8

Опубликована: Янв. 25, 2021

Язык: Английский

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SLC45A2 protein stability and regulation of melanosome pH determine melanocyte pigmentation

Molecular Biology of the Cell, Год журнала: 2020, Номер 31(24), С. 2687 - 2702

Опубликована: Сен. 23, 2020

SLC45A2 encodes a putative transporter expressed primarily in pigment cells. mutations cause oculocutaneous albinism type 4 (OCA4) and polymorphisms are associated with pigmentation variation, but the localization, function, regulation of its variants remain unknown. We show that localizes to cohort mature melanosomes only partially overlaps expressing chloride channel OCA2. ectopically HeLa cells lysosomes raises lysosomal pH, suggesting melanocytes expression, like OCA2 results deacidification maturing support melanin synthesis. Interestingly, overexpression compensates for loss expression pigmentation. Analyses SLC45A2- OCA2-deficient mouse likely functions later during melanosome maturation than Moreover, light skin-associated allelic F374 variant restores moderate SLC45A2-deficient due rapid proteasome-dependent degradation resulting lower protein levels dark L374 variant. Our data suggest maintains neutralization is initially orchestrated by transient activity melanization at late stages maturation, common imparts reduced instability.

Язык: Английский

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Identification and functional evaluation of GRIA1 missense and truncation variants in individuals with ID: An emerging neurodevelopmental syndrome

The American Journal of Human Genetics, Год журнала: 2022, Номер 109(7), С. 1217 - 1241

Опубликована: Июнь 7, 2022

GRIA1 encodes the GluA1 subunit of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors, which are ligand-gated ion channels that act as excitatory receptors for neurotransmitter L-glutamate (Glu). AMPA (AMPARs) homo- or heteromeric protein complexes with four subunits, each encoded by different genes, to GRIA4. Although GluA1-containing AMPARs have a crucial role in brain function, human phenotype associated deleterious sequence variants has not been established. Subjects de novo missense and nonsense were identified through international collaboration. Detailed phenotypic genetic assessments subjects carried out pathogenicity was evaluated vitro characterize changes AMPAR function expression. In addition, two Xenopus gria1 CRISPR-Cas9 F0 models established vivo consequences. Seven unrelated individuals rare identified. One individual homozygous variant (p.Arg377Ter), six had heterozygous variations (p.Arg345Gln, p.Ala636Thr, p.Ile627Thr, p.Gly745Asp), p.Ala636Thr recurrent three individuals. The cohort revealed neurodevelopmental disorder mostly affecting cognition speech. Functional evaluation major subtypes carrying mutations showed profoundly perturb receptor function. stop-gain completely destroys expression AMPARs. show transient motor deficits, an intermittent seizure phenotype, significant impairment working memory mutants. These data support developmental caused both

Язык: Английский

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Autism gene variants disrupt enteric neuron migration and cause gastrointestinal dysmotility

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Март 6, 2025

The co-occurrence of autism and gastrointestinal distress is well-established, yet the molecular underpinnings remain unknown. identification high-confidence, large-effect genes offers opportunity to identify convergent, underlying biology by studying these in context system. Here we show that expression enriched human prenatal gut neurons their migratory progenitors, suggesting development and/or function may be disrupted autism-associated genetic variants, leading dysfunction. document prevalence issues patients with variants sixteen genes, highlighting dysmotility, consistent potential enteric neuron Using Xenopus tropicalis, individually target five (SYNGAP1, CHD8, SCN2A, CHD2, DYRK1A) observe neuronal progenitor migration for each. Further analysis DYRK1A reveals perturbation causes dysmotility vivo, which can ameliorated treatment either two serotonin signaling modulators, identified vivo drug screening. This work suggests atypical contributes commonly seen individuals a productive therapeutic pathway.

Язык: Английский

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Creating cell and animal models of human disease by genome editing using CRISPR/Cas9

The Journal of Gene Medicine, Год журнала: 2019, Номер 21(4)

Опубликована: Фев. 20, 2019

A set of unique sequences in bacterial genomes, responsible for protecting bacteria against bacteriophages, has recently been used the genetic manipulation specific points genome. These systems consist one RNA component and enzyme component, known as CRISPR ("clustered regularly interspaced short palindromic repeats") Cas9, respectively. The present review focuses on applications CRISPR/Cas9 technology development cellular animal models human disease. Making a desired alteration depends design molecules that guide endonucleases to favorable genomic location. With discovery technology, researchers are able achieve higher levels accuracy because its advantages over alternative methods editing genome, including simple design, high targeting efficiency ability create simultaneous alterations multiple sequences. factors allow apply this creating diseases by knock-in, knock-out Indel mutation strategies, such Huntington's disease, cardiovascular disorders cancers. Optimized will facilitate access valuable novel with respect innovative drug gene therapy.

Язык: Английский

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Xenopus Resources: Transgenic, Inbred and Mutant Animals, Training Opportunities, and Web-Based Support

Frontiers in Physiology, Год журнала: 2019, Номер 10

Опубликована: Апрель 25, 2019

Two species of the clawed frog family, Xenopus laevis and tropicalis, are widely used as tools to investigate both normal disease-state biochemistry, genetics, cell biology developmental biology. To support specialist non-specialist scientists needing access these models for their research, a number centralised resources exist around world. These include centres that hold live frozen stocks transgenic, inbred mutant animals molecular resources. This infrastructure is supported by model organism database. Here we describe much this encourage community make best use it guide resource in developing new lines libraries.

Язык: Английский

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Xenopus leads the way: Frogs as a pioneering model to understand the human brain

genesis, Год журнала: 2020, Номер 59(1-2)

Опубликована: Дек. 27, 2020

From its long history in the field of embryology to recent advances genetics, Xenopus has been an indispensable model for understanding human brain. Foundational studies that gave us our first insights into major embryonic patterning events serve as a crucial backdrop newer avenues investigation organogenesis and organ function. The vast array tools available laevis tropicalis allows interrogation developmental phenomena at all levels, from molecular behavioral, application CRISPR technology enabled disorder risk genes higher-throughput manner. As only tetrapod which stages are easily manipulated observed, frogs provide unique opportunity study development earliest stages. All these features make premier studying brain, notoriously complex process demands fertilization beyond. Importantly, core processes brain conserved between human, underlining advantages this model. This review begins by summarizing discoveries made amphibians form cornerstones vertebrate neurodevelopmental biology goes on discuss have catapulted relation disease. we engage new era patient-driven gene discovery, offers exceptional potential uncover underlying disorders move towards rational drug design.

Язык: Английский

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Maximizing CRISPR/Cas9 phenotype penetrance applying predictive modeling of editing outcomes in Xenopus and zebrafish embryos

Scientific Reports, Год журнала: 2020, Номер 10(1)

Опубликована: Сен. 4, 2020

Abstract CRISPR/Cas9 genome editing has revolutionized functional genomics in vertebrates. However, edited F 0 animals too often demonstrate variable phenotypic penetrance due to the mosaic nature of outcomes after double strand break (DSB) repair. Even with high efficiency levels editing, phenotypes may be obscured by proportional presence in-frame mutations that still produce protein. Recently, studies cell culture systems have shown CRISPR/Cas9-mediated can dependent on local sequence context and predicted computational methods. Here, we similar approaches used forecast gene Xenopus tropicalis , laevis, zebrafish. We show a publicly available neural network previously trained mouse embryonic stem cultures (InDelphi-mESC) is able accurately predict early vertebrate embryos. Our observations direct implications for experiment design, allowing selection guide RNAs repair outcome signatures enriched towards frameshift mutations, maximization phenotype generation.

Язык: Английский

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CRISPR/Cas9 disease models in zebrafish and Xenopus: The genetic renaissance of fish and frogs

Drug Discovery Today Technologies, Год журнала: 2018, Номер 28, С. 41 - 52

Опубликована: Июль 31, 2018

Язык: Английский

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Neurodevelopmental disorder risk gene DYRK1A is required for ciliogenesis and brain size in Xenopus embryos

Development, Год журнала: 2020, Номер unknown

Опубликована: Янв. 1, 2020

DYRK1A (dual specificity tyrosine-(Y)-phosphorylation-regulated kinase 1 A) is a high confidence autism risk gene that encodes conserved kinase. In addition to autism, patients with putative loss of function variants in exhibit microcephaly, intellectual disability, developmental delay, and/or congenital anomalies the kidney and urinary tract. also located within critical region for Down syndrome; therefore, understanding role brain development crucial pathobiology multiple disorders. To characterize this gene, we used diploid frog, Xenopus tropicalis. We discover Dyrk1a expressed ciliated tissues, localizes ciliary axonemes basal bodies, required ciliogenesis. demonstrate mitotic spindles its inhibition leads decreased forebrain size, abnormal cell cycle progression, death during development. These findings provide hypotheses about potential mechanisms underscore utility X. tropicalis as model system neurodevelopmental

Язык: Английский

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