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The application of CRISPR-Cas9 for the creation of multiple mutants in the genes coding for the nascent polypeptide associated complex

Public support

  • Provider

    Ministry of Education, Youth and Sports

  • Programme

    INTER-EXCELLENCE

  • Call for proposals

    SMSM2020LTC01

  • Main participants

    Ústav experimentální botaniky AV ČR, v. v. i.

  • Contest type

    VS - Public tender

  • Contract ID

    MSMT-21080/2020-24

Alternative language

  • Project name in Czech

    The application of CRISPR-Cas9 for the creation of multiple mutants in the genes coding for the nascent polypeptide associated complex

  • Annotation in Czech

    Flower and silique developments represent important processes of plant reproduction. Conventional Arabidopsis thaliana double homozygous mutant in the genes coding for the β-subunit of the nascent polypeptide associated complex (that were acquired by the cross of the available T-DNA insertion lines) showed notable phenotypic defects in the flowers and siliques. Moreover, seeds of the double homozygous mutant showed a reduced germination efficiency under the salt and osmotic stress. The aim of this project is to employ modern genome editing techniques to acquire the multiple mutants in the genes coding for the subunits of the nascent polypeptide-associated complex (a quintuple mutant of genes encoding its α-subunit and a double mutant in genes encoding the β-subunit). These genome-edited plants will be characterized in more detail and will elucidate the role of nascent polypeptide-associated complex during flower and silique developments of Arabidopsis thaliana. The key results of this project will include (1) the creation of various double mutants in the β-subunit of nascent polypeptide-associated complex and various quintuple mutants in the α-subunit of nascent polypeptide-associated complex by the modern genome-editing techniques (CRISPR-Cas9), (2) the characterization of various genome-editing events and phenotypic characterization of the plants with edited genomic DNA of the genes encoding the nascent polypeptide-associated complex, (3) the comparison of the genome-edited plants with the conventional double homozygous mutants of the β-subunit acquired by the cross of the available T-DNA insertion lines, (4) the creation of the plants with organ-specific genome editing by CRISPR-TSKO.

Scientific branches

  • R&D category

    ZV - Basic research

  • OECD FORD - main branch

    10611 - Plant sciences, botany

  • OECD FORD - secondary branch

  • OECD FORD - another secondary branch

  • CEP - equivalent branches <br>(according to the <a href="http://www.vyzkum.cz/storage/att/E6EF7938F0E854BAE520AC119FB22E8D/Prevodnik_oboru_Frascati.pdf">converter</a>)

    EF - Botany

Completed project evaluation

  • Provider evaluation

    V - Vynikající výsledky projektu (s mezinárodním významem atd.)

  • Project results evaluation

    The aim of the project was to create a quintuple mutant in genes encoding for the α-subunit of the nascent polypeptide-associated complex (NAC) and a double mutant in genes encoding for NACβ by CRISPR-Cas9 technology. Both mutants were notably delayed in their development, and the nacα mutants showed also less chlorophyll in their leaves but the other photosynthetic parameters were not influenced.

Solution timeline

  • Realization period - beginning

    Jun 1, 2020

  • Realization period - end

    Mar 31, 2023

  • Project status

    U - Finished project

  • Latest support payment

    Feb 17, 2023

Data delivery to CEP

  • Confidentiality

    S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

  • Data delivery code

    CEP24-MSM-LT-U

  • Data delivery date

    Jul 1, 2024

Finance

  • Total approved costs

    4,863 thou. CZK

  • Public financial support

    4,863 thou. CZK

  • Other public sources

    0 thou. CZK

  • Non public and foreign sources

    0 thou. CZK