![]() ![]() ![]() RT-PCR and RNA-Seq analysis showed that the melanin PKS cluster was down-regulated in infected banana as compared to growth in culture. fijiensis PKS sequences, but three others were not found in either species. eumusae, showed that these two species have close homologs to five of the M. fijiensis PKS sequences with those of two other banana pathogens, M. One of the PKS sequences was not similar (< 60% similarity) to sequences in any of the 103 genomes, suggesting that it encodes a unique compound. A search for homologs among available genomic sequences from 103 Dothideomycetes identified close homologs (>80% similarity) for six of the PKS sequences. None of the other clusters were closely aligned with genes encoding known polyketides, however three of the PKS genes fell into clades with clusters encoding alternapyrone, fumonisin, and solanapyrone produced by Alternaria and Fusarium species. Phylogenetic analysis identified a putative PKS cluster encoding melanin biosynthesis. Gene clusters contained types of genes frequently found in PKS clusters including genes encoding transporters, oxidoreductases, methyltransferases, and non-ribosomal peptide synthases. Analysis of the PKS domains identified three of the PKS enzymes as non-reducing and two as highly reducing. fijiensis into the 23rd percentile for the number of PKS genes compared to other Dothideomycetes. Eight PKS gene clusters were identified in the M. fijiensis genome sequence to predict PKS genes and their gene clusters and make bioinformatics predictions about the types of compounds produced by these clusters. By excellent research training of 8 ESRs on a scientifically and economically highly relevant topic and additional training in transferable skills, IMGENE will educate novel leaders with great career perspectives in industry and academia.Mycosphaerella fijiensis, causal agent of black Sigatoka disease of banana, is a Dothideomycete fungus closely related to fungi that produce polyketides important for plant pathogenicity. In addition, IMGENE addresses crucial ethical questions related to the application of genome editing technology in animals, plants, and humans, which have to be solved to gain acceptance by the society. Combining complementary knowledge on protein chemistry, molecular biology, cellular biology, viral vectors, transgenic mice, gene therapy, and bioinformatics present in the network, IMGENE will establish novel tools and protocols for improved CRISPR genome editing efficiency that will be of immediate benefit for health and life science research, the pharmaceutical industry, and the application of gene therapy. The IMGENE consortium consisting of 6 academic and 1 industrial beneficiary (AstraZeneca), 2 industrial partners (Taconic MilliporeSigma), and the patient organization Genetic Alliance, aims to improve the genome editing efficiency of CRISPR by research training of 8 ESRs to unleash the full potential of this technology. To our best knowledge, IMGENE is the only concerted approach to tackle the important problem of low genome editing efficiency of CRISPR in a multidisciplinary, intersectorial manner. IMGENE unites expert European research groups of academia and industry to address by innovative and complementary approaches the low efficiency of precise genome editing using CRISPR technology. However, the efficiency of introducing defined changes into the genome by CRISPR is still low, currently limiting its application in basic research, industry and gene therapy. ![]() CRISPR genome editing technology is considered to become the greatest technological improvement in biomedical research since the invention of the polymerase chain reaction 25 years ago and pharmaceutical companies as well as academic research are eager to apply it. ![]()
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