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HD-DittoGraph SIGNED

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

Total Cost €


EC-Contrib. €






 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.

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

Project "HD-DittoGraph" data sheet

The following table provides information about the project.


Organization address
address: Via Festa Del Perdono 7
city: MILANO
postcode: 20122

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


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


 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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