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HD-DittoGraph SIGNED

HD-DittoGraph: a digital human Embryonic Stem Cell platform for Huntington's repeats

Total Cost €

0

EC-Contrib. €

0

Partnership

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 HD-DittoGraph project word cloud

Explore the words cloud of the HD-DittoGraph project. It provides you a very rough idea of what is the project "HD-DittoGraph" about.

intend    gain    postmitotic    huntington    causative    perform    cis    contribution    screenings    construction    mitotic    genetic    genome    htt    sequences    modifiers    subsequently    association    introduce    relies    computational    cells    efficient    trans    novelty    striatal    embryonic    platform    pipelines    divisions    size    platforms    identification    barcodes    transcriptome    networks    locus    pure    donor    hes    disease    libraries    sequence    length    generating    neuronal    human    shaped    crispr    modifier    functional    validated    gene    mechanisms    cag    exon    cell    dividing    cas9    inducible    individually    instability    tested    couples    library    screen    act    multiple    constructs    stem    simultaneous    neurons    acting    differentiating    flanking    huntingtin    generation    unequivocally    gof    candidate    corresponding    hoc    neurogenic    barcoded    sgrnas    lof    causing    insights    thousands    ad    genes    sequencing    molecular    function    repeat    expansion    unstable   

Project "HD-DittoGraph" data sheet

The following table provides information about the project.

Coordinator
UNIVERSITA DEGLI STUDI DI MILANO 

Organization address
address: Via Festa Del Perdono 7
city: MILANO
postcode: 20122
website: www.unimi.it

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country Italy [IT]
 Total cost 2˙040˙943 €
 EC max contribution 2˙040˙943 € (100%)
 Programme 1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
 Code Call ERC-2016-ADG
 Funding Scheme ERC-ADG
 Starting year 2018
 Duration (year-month-day) from 2018-03-01   to  2023-02-28

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    UNIVERSITA DEGLI STUDI DI MILANO IT (MILANO) coordinator 1˙868˙443.00
2    FONDAZIONE ISTITUTO NAZIONALE DI GENETICA MOLECOLARE INGM IT (MILANO) participant 172˙500.00

Map

 Project objective

This proposal is aimed at identifying the molecular mechanisms that have brought the human Huntington Disease-causing Huntingtin (Htt) exon 1, with its pure and unstable CAG repeat, to be shaped the way it is today. Specifically, we intend to screen for genetic elements affecting Htt repeat length instability in dividing and postmitotic neuronal cells. The novelty of our approach relies on the construction of a human embryonic stem (hES) cell platform that couples highly efficient CRISPR/Cas9 technology with genome-wide screenings and third generation sequencing, to test the contribution of thousands of unequivocally barcoded cis and trans modifiers on Htt exon 1 repeats instability.

In Aim 1, we will test the contribution of cis-modifiers to repeat instability during multiple mitotic divisions, by generating a hES cell platform where we will subsequently introduce a barcoded donor library of different Htt exon 1 constructs, with different CAG and flanking sequences, at the Htt locus. In Aim 2 our hES cell platform will be implemented with inducible Cas9 elements and sgRNAs libraries to perform genome-wide loss and gain of function (LOF, GOF) screenings of trans-acting modifiers of repeat sequence and size. The sgRNAs will act as barcodes for the modifier genes, allowing to test their causative role on repeat size changes. In Aim 3, we will exploit the neurogenic potential of hES cells in our LOF and GOF platforms to identify Htt exon 1 repeat modifiers in differentiating striatal neurons. Candidate modifier genes will be individually validated and tested for their functional impact on gene networks by transcriptome analysis. In all approaches, third generation sequencing and ad hoc computational pipelines will allow the simultaneous identification of the repeat changes and their association to the corresponding modifiers. Overall, this research proposal is expected to provide key molecular and genetic insights into the process of Htt repeat expansion in human

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