CRISPR Laboratorium

    Facilitet: Laboratorie

    • Placering

      Danmark

    Beskrivelse

    The CRISPR/Cas9 system enables precise and efficient targeted genome engineering. It is a two-component system which uses a single guide RNA sequence (sgRNA) to direct the Cas9 endonuclease to complementary sequence regions in the genome, leading to the generation of double-stranded DNA breaks. Originally, the CRISPR/Cas9 system was part of the bacterial immune response towards virus and bacteriophage infections but with the transfer into eukaryotic cells the CRISPR/Cas9 induced DNA breaks strongly activates the DNA damage repair pathways of non-homologous end joining (NHEJ) and homology-directed repair (HDR). When the breaks are repaired by NHEJ, error-prone base insertions and deletions result in frequent frameshift mutations and premature stop codons eliminating the production of functional protein. In the presence of a homologous repair template double-stranded DNA breaks can via HDR be repaired substituting precise sequences. With optimized sgRNA sequences it is possible to target all genes in the human genome and combining systematic genetic screening using lentiviral CRISPR/Cas9 technology and an external selection strategy a link between geno- and phenotype can be generated.

    The hematological CRISPR research project have a special interest in identify genes that alone or in combination control development of drug resistance and differentiation in hematopoietic cancers. Using CRISPR/Cas9 based genome editing, hematopoietic cells will be challenged in drug exposure screens and cellular differentiation assays aiming at identifying the genes that directly affect drug response and cellular maturation.

    We anticipate that genetic aberrations identified in in vitro CRISPR/Cas9 analysis holds valuable clinical outcome related information. For selected genetic aberrations we expect important biological mechanisms of intrinsic and acquired treatment resistance and cellular differentiation to be determined in functional assays and we hope to identify novel active drug combinations able to reverse treatment resistance identified.


    The CRISPR lab is located at Forskningens Hus, Sdr Skovvej 15, Rumnummer 15.06.124c.
    It is classified as gene-technology class 2, due to the usage of HIV-1 derived vector systems.
    Projects must prior to initiation be reported to and obtain permission from the Danish Working Environment Authority.

    Fingeraftryk

    Clustered Regularly Interspaced Short Palindromic Repeats
    Guide RNA
    Double-Stranded DNA Breaks
    Genes
    Genome
    Technology
    Frameshift Mutation
    DNA Breaks
    Nonsense Codon
    Endonucleases
    Genetic Testing
    Eukaryotic Cells
    Human Genome
    Virus Diseases
    Drug Combinations
    Drug Resistance
    DNA Repair
    Pharmaceutical Preparations
    Bacteriophages
    DNA Damage