Genetic mosaicism, a condition in which an organ includes cells with different genotypes, is frequently present in monogenic diseases of the central nervous system caused by the random inactivation of the X-chromosome, in the case of X-linked pathologies, or by somatic mutations affecting a subset of neurons. The comprehension of the mechanisms of these diseases and of the cell-autonomous effects of specific mutations requires the generation of sparse mosaic models, in which the genotype of each neuron is univocally identified by the expression of a fluorescent protein in vivo. Here, we show a dual-color reporter system that, when expressed in a floxed mouse line for a target gene, leads to the creation of mosaics with tunable degree. We demonstrate the generation of a knockout mosaic of the autism/epilepsy related gene PTEN in which the genotype of each neuron is reliably identified, and the neuronal phenotype is accurately characterized by two-photon microscopy.

Modelling genetic mosaicism of neurodevelopmental disorders in vivo by a Cre-amplifying fluorescent reporter

Nardi, Gabriele;
2020

Abstract

Genetic mosaicism, a condition in which an organ includes cells with different genotypes, is frequently present in monogenic diseases of the central nervous system caused by the random inactivation of the X-chromosome, in the case of X-linked pathologies, or by somatic mutations affecting a subset of neurons. The comprehension of the mechanisms of these diseases and of the cell-autonomous effects of specific mutations requires the generation of sparse mosaic models, in which the genotype of each neuron is univocally identified by the expression of a fluorescent protein in vivo. Here, we show a dual-color reporter system that, when expressed in a floxed mouse line for a target gene, leads to the creation of mosaics with tunable degree. We demonstrate the generation of a knockout mosaic of the autism/epilepsy related gene PTEN in which the genotype of each neuron is reliably identified, and the neuronal phenotype is accurately characterized by two-photon microscopy.
2020
Settore BIO/11 - Biologia Molecolare
Settore BIO/09 - Fisiologia
Action potentials; animals; newborn; disease models, Animal; electroencephalography; fluorescent dyes; gene expression; genes, reporter; HEK293 cells; humans; integrases; mice; inbred C57BL; mosaicism; neurodevelopmental disorders; NIH 3T3 cells; PTEN phosphohydrolase; tamoxifen; focal cortical dysplasias; somatic mutations; diagnostic methods; mouse model; spectrum; brain; pten; recombination; mtor; malformations
   Early dysfunctions of intercellular signalling in brain disorders
   MUR
   PRIN-2017

   Intracellular chloride dynamics in autistic brain: a better understanding is needed for tailored cures
   Fondazione Telethon
   GP19281
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11384/136802
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