C. elegans as a model to study PTEN’s regulation and function

Liu, Jun; Chin-Sang, Ian D.

doi:10.1016/j.ymeth.2014.12.009

Cited by 18 publications

(18 citation statements)

References 101 publications

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“…Since ASD is a behaviorally diagnosed disorder, and because deficits in sensory processing are a core feature of ASD present in >95% of cases 58 , we tested the effects of PTEN variants on sensorimotor neural circuit function in an intact animal model. We used a behavioral assay in C. elegans as they have a unique advantage of being the only animal known to survive homozygous null mutations in PTEN, allowing us to directly assess the functional consequences of PTEN variants in a transgenic rescue assay in vivo 52 . C. elegans exhibit robust navigation up a concentration gradient toward a point source of NaCl.…”

Section: Pten Variant Effects On Sensory Processing and Behavior In Cmentioning

confidence: 99%

Multi-model functionalization of disease-associated PTEN missense mutations identifies multiple molecular mechanisms underlying protein dysfunction

Post

Belmadani

Ganguly

et al. 2019

Preprint

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ABSTRACTFunctional variomics provides the foundation for personalized medicine by linking genetic variation to disease expression, outcome and treatment, yet its utility is dependent on appropriate assays to evaluate mutation impact on protein function. To fully assess the effects of 106 missense and nonsense variants of PTEN associated with autism spectrum disorder, somatic cancer and PHTS, we take a deep phenotypic profiling approach using 18 assays in 5 model systems spanning diverse cellular environments ranging from molecular function to neuronal morphogenesis and behavior. Variants inducing instability occurred across the protein, resulting in partial to complete loss of function (LoF), which was well correlated across models. However, assays were selectively sensitive to variants located in substrate binding and catalytic domains, which exhibited complete LoF or dominant negativity independent of effects on stability. Our results indicate that full characterization of variant impact requires assays sensitive to instability and a range of protein functions.

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Section: Pten Variant Effects On Sensory Processing and Behavior In Cmentioning

confidence: 99%

Multi-model functionalization of disease-associated PTEN missense mutations identifies multiple molecular mechanisms underlying protein dysfunction

Post

Belmadani

Ganguly

et al. 2019

Preprint

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“…2) C. elegans has a single PTEN 147 ortholog called daf-18 ( Fig. 2 A-D) and transgenic overexpression of human PTEN using 148 extrachromosomal arrays has been shown to rescue reduced longevity and dauer 149 defective phenotypes induced by mutations in daf-18 (Liu and Chin-Sang, 2015;Solari et 150 al., 2005). 3) C. elegans harboring homozygous daf-18 null alleles are viable and display 151 superficially normal morphology and spontaneous locomoter behavior (Mihaylova et al 152 1999 and Fig.…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Cas9 human gene replacement and phenomic characterization inCaenorhabditis elegansto understand the functional conservation of human genes and decipher variants of uncertain significance

McDiarmid

Loewen

et al. 2018

Preprint

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Our ability to sequence genomes has vastly surpassed our ability to interpret the genetic variation we discover. This presents a major challenge in the clinical setting, where the recent application of whole exome and whole genome sequencing has uncovered thousands of genetic variants of uncertain significance. Here, we present a strategy for targeted human gene replacement and phenomic characterization based on CRISPR-Cas9 genome engineering in the genetic model organism Caenorhabditis elegans that will facilitate assessment of the functional conservation of human genes and structure-function analysis of disease-associated variants with unprecedented precision.We validate our strategy by demonstrating that direct single-copy replacement of the C. elegans ortholog (daf-18) with the critical human disease-associated gene Phosphatase and Tensin Homolog (PTEN) is sufficient to rescue multiple phenotypic abnormalities caused by complete deletion of daf-18, including complex chemosensory and mechanosenory impairments. In addition, we used our strategy to generate animals harboring a single copy of the known pathogenic lipid phosphatase inactive PTEN variant (PTEN-G129E) and showed that our automated in vivo phenotypic assays could accurately and efficiently classify this missense variant as loss-of-function. The integrated nature of the human transgenes allows for analysis of both homozygous and heterozygous variants and greatly facilitates high-throughput precision medicine drug screens. By combining genome engineering with rapid and automated phenotypic characterization, our strategy streamlines identification of novel conserved gene functions in complex sensory and learning phenotypes that can be used as in vivo functional assays to decipher variants of uncertain significance. remarkably efficient and robust in C. elegans (Dickinson and Goldstein, 2016; Norris et 70 al., 2015). 71Here, we present a broadly applicable strategy that adapts CRISPR-Cas9 genome 72 engineering for targeted replacement of C. elegans genes with human genes. We illustrate 73 how the library of knockout and humanized transgenics generated with this approach can 74 be efficiently combined with automated machine vision phenotyping to rapidly discover 75 novel gene functions, assess the functional conservation of human genes, and how this 76 will allow for analysis of the effects of variants of uncertain significance with 77 unprecedented precision. It is our hope that the human gene replacement and phenomic 78 characterization strategy delineated in this article will serve both basic and health 79 researchers alike, by serving as an open and shareable resource that will aid any genome 80 engineer interested in understanding the functional conservation of human genes, and the 81 functional consequences of their variants. 82 83 Results 84A general genome editing strategy for direct replacement of a C. elegans gene with a 85 single copy of its human ortholog 86To replace the Open Reading Frame (ORF) of an orthologous gene with a human 87 gene ...

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“…Moreover, the role of DAF-18 with other aging related signaling components of C. elegans further supports a potential role of PTEN in mammalian aging (J. Liu & Chin-Sang, 2015;Solari et al, 2005).…”

Section: Drosophila Melanogaster One Of the Most Important Genetic Mmentioning

confidence: 58%

“…Studies have shown that human PTEN can substitute for DAF-18 and rescue the longevity defects caused due to DAF-18 mutation (J. Liu & Chin-Sang, 2015;Solari et al, 2005). Homozygous deletion of daf-18 in C. elegans results in a shorter life span but results in viable and fertile worms.…”

Section: Drosophila Melanogaster One Of the Most Important Genetic Mmentioning

confidence: 99%

“…However, it has also been shown to be involved in a variety of other functions, including regulation of cell size, metabolism, aging and development of neurons (J. Liu & Chin-Sang, 2015). Orthologs of PTEN have been identified in Drosophila melanogaster (Huang et al, 1999b), Caenorhabditis elegans (Gil, Malone Link, Liu, Johnson, & Lees, 1999;Solari et al, 2005), and budding yeast (Heymont et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

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Exploring the role of PTN1 in ustilago maydis.

Pillai¹,

Krishna²

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Signal transduction is a key aspect of biological life. PTEN (phosphatase and tensin homolog) is a widely studied signaling protein in mammalian systems, where it acts as a tumor suppressor. Numerous studies have conclusively demonstrated the anti-cancerous effect of this protein. Mutations in the PTEN gene have been implicated in several disorders including cancer, diabetes, and Cowden syndrome, among others. It is part of the PI3K-PTEN-AKT-mTOR signaling pathway and is responsible for various metabolic and cellular activities. This pathway is found to be conserved across several eukaryotic systems including yeast, Drosophila melanogaster and Caenorhabditis elegans. While PTEN orthologues have been characterized in ascomycete fungi, it has not been reported in any basidiomycete fungi. The initial part of the current study was aimed at the characterization of PTEN orthologue, ptn1, of Ustilago maydis, a basidiomycete fungus. When the U. maydis, ptn1 gene was either deleted or overexpressed, it was found that the deletion of this gene lead to reduced virulence, spore production, and germination rate; effects of the overexpression were more subtle or not discernable. Previous studies have indicated that two other signaling proteins, Rho1 and Pdc1, are physical interactors of Ptn1. The goal of the latter part of the present study was to explore the functional interaction of Ptn1 with vi Rho1 and Pdc1. Our experiments indicated a functional connection between these genes, especially in processes that are related to stress tolerance, cellular morphology, and virulence of the fungus, suggesting that they may be elements of the same regulatory pathway. We predict that the pdc1 interaction with ptn1 may be the reverse of that of rho1 with ptn1. In addition, our experiments with an orthologue of S6K1, aga1, showed that its deletion leads to some of the same phenotypes seen with that of ptn1, pdc1 and/or rho1. The current phenotypes of these mutant strains when considered in the context of already available information about the role of these genes in other systems indicate that they may be part of a pathway that modulates cytoskeletal rearrangements. vii

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C. elegans as a model to study PTEN’s regulation and function

Cited by 18 publications

References 101 publications

Multi-model functionalization of disease-associated PTEN missense mutations identifies multiple molecular mechanisms underlying protein dysfunction

Multi-model functionalization of disease-associated PTEN missense mutations identifies multiple molecular mechanisms underlying protein dysfunction

CRISPR-Cas9 human gene replacement and phenomic characterization inCaenorhabditis elegansto understand the functional conservation of human genes and decipher variants of uncertain significance

Exploring the role of PTN1 in ustilago maydis.

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