Browsing by Author "Aksoy, Emre"

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    General guidelines for CRISPR/Cas-based genome editing in plants
    (2022) Aksoy, Emre; Yildirim, Kubilay; Kavas, Musa; Kayihan, Ceyhun; Yerlikaya, Bayram Ali; Calik, Irmak; Sevgen, Ilkay; Demirel, Ufuk; 36107373
    CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) technology is a versatile genome editing tool that has been used to improve agriculturally important plant traits. Due to its precision, CRISPR/Cas9 is more effective than either conventional plant breeding methods or standard genetic engineering approaches for the rapid development of new varieties resilient to climate change. In addition to knowledge in tissue culture-based plant transformation, effective gene-specific single guide RNA (sgRNA) design, prediction of its off-target effect and utilization of vectors, promoters, Cas proteins and terminators is required for CRISPR/Cas9. Various bioinformatics tools are available for the best sgRNA design and screening of the off-targets. Various tools are used in the delivery of CRISPR/Cas components into cells and the genome. Moreover, some recent studies proved the simultaneous silencing of different paralogs in the same family or several genes working in the same pathway by using multiple-target sgRNA designs. This review summarizes the type of promoters, Cas proteins, recognition sequences, and terminators available for the development of knock-out and overexpression plant lines. It also provides a general guideline for the development of genome-edited plants from the design of sgRNAs to the selection of non-transgenic genome-edited T-2 generation.
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    Identification and expression profiling of toxic boron-responsive microRNAs and their targets in sensitive and tolerant wheat cultivars
    (2021) Kayihan, Doga Selin; Aksoy, Emre; Kayihan, Ceyhun; 0000-0003-1684-4147; AAW-8352-2021
    Post transcriptional gene regulation in wheat cultivars caused by boron (B) toxicity has not been reported to date. In this study, two wheat cultivars Bolal-2973 (B-tolerant) and Atay-85 (B-sensitive) were compared with regard to the transcriptional regulation of miR319, miR172 and miR398 and their plausible target genes in order to address differences in their performances under high B levels. The expression levels of Cu/ZnSOD gene were found to be greater in Atay exposed to toxic B compared to Bolal, as verified by stable expression level of miR398. In wheat cultivars, both toxic B concentrations might cause an induction of leaf senescence mechanism due to stable level of JA and ethylene-related miRNAs, miR172 and miR319. miR172 targeting transcription factor TOE1 was only induced under B toxicity in sensitive cultivar Atay. However, MY133 as target of miR319 was significantly upregulated under toxic B in both cultivars. Also, computational prediction of these miRNA targets in wheat was performed and their orthologs in Arabidopsis thaliana were determined. Functional protein association networks of proteins encoded by miRNA targets and gene ontology enrichment analyses of these genes were identified. We identified new sets of genes that are targets of miR172, miR319 and miR398 in T. aestivum. In addition, miR172, miR319 and miR398 are responsive to different nutrient deficiencies or toxicities such as Fe, P, B, S and Cu, suggesting crosstalk between the post-transcriptional regulatory mechanisms involving miRNAs in plants.