TAILIEUCHUNG - Genome editing of rice PFT1 gene to study its role in rice sheath blight disease resistance

The CRISPR/Cas9 system has been used predominantly for precise editing of the plant genomes. In the present study, we have made an application of CRISPR/Cas9 system for targeted mutagenesis in rice(Oryza sativa L.) targeting the Phytochrome and Flowering Time 1 (PFT1) gene known to be involved in disease susceptibility in Arabidopsis for rootinfecting hemibiotrophic fungal pathogen Fusarium oxysporum. Hence, to confirm the role of OsPFT1 gene in rice sheath blight disease, CRISPR-Cas9 gene editing tool is being employed for generating OsPFT1 gene knock out in rice. Two guide RNAs (gRNAs) were designed to pair with distinct OsPFT1 gene region followed by protospacer-adjacent motif (PAM). Rice genome editing constructs were cloned and mobilized into the Agrobacterium strain LBA4404. The transgene (Cas9-gRNA) introduction into indica rice variety ASD16was done via Agrobacterium-mediated rice transformation by using immature embryos as explants. The gRNA will direct the Cas9 nuclease to generate double-strand breaks (DSB) at the specific sites of OsPFT1 gene, thereby introducing mutations at the DSB by error-prone non-homologues end joining repair mechanism. Through PCR analysis, the presence of the hpt transgene was identified. Homozygous gene-edited transgenic rice plants will be identified and further will be subjected to sheath blight disease screening. | Genome editing of rice PFT1 gene to study its role in rice sheath blight disease resistance

• Nông nghiệp
• Gene editing
• CRISPR Cas9
• Rice transformation
• Phytochrome and flowering time 1
• Sheath blight resistance
• Plant virus
• RNA silencing
• Genome editing
• Antiviral breeding
• Gene silencing to genome editing
• BMC Biotechnology
• High oleic acid
• ahFAD2 gene
• Anti oxidation
• Genome Biology
• PAM stringency
• Off targeting
• C containing PAM
• Multiplex gene editing
• tRNA processing
• Nicotiana tabacum
• Oryza sativa
• Non coding RNAs
• Base editing
• Exon skipping
• Alternative splicing
• Synthetic biology
• BMC Genomics
• Genomic complexity
• Sequencing technology development
• Complex genetics
• Precise genetic editing
• Genome engineering
• Fluorescence activated cell sorting
• Mutation enrichment
• Nicotiana benthamiana
• Migratory locust
• Pre mRNA splicing
• BMC Plant Biology
• Multiple gene mutations
• Assembly of multiple gRNAs
• Transfer DNA
• Plant Biology
• Pepper genome editing
• Capsicum annuum
• C annuum Dempsey
• Pepper leaf protoplasts
• Pepper callus protoplasts
• Betula platyphylla
• White birch
• Chloroplast genome
• RNA editing
• Simple sequence repeat
• Next generation sequencing
• Translocation detection
• Journal of Biology
• CRISPR Cas9 technology
• Bioethical issues
• Animal models
• Ototoxic deafness
• Aminoglycoside antibiotics
• Hair cell
• Anti apoptosis
• RNA sequences
• Thereby increasing protein diversity
• Gene regulation
• Cancer related immune pathways
• Cajal body
• Cancer mutations
• CRISPR Cas9 genome editing
• Facilitate indefinite proliferation
• Stem cells
• Bacterial type II CRISPR Cas9 system
• Genome editing experiments
• CRISPR associated protein 9
• Single guide RNA
• Cell stress
• Innate immune cells
• Gene knockdown
• Viral suppressor
• CRISPR/Cas9 system
• Essential gene
• Multicopy gene
• Foreign DNA
• Bait DNA
• Xenogeneic silencers
• Nucleoid associated proteins
• Intrachromosomal homologous recombination
• Gene targeting
• Translational recoding
• Abolish gene function
• Definitive null
• Knockout first
• Gene disruption
• Exon skipping
• Disrupt genes
• Histone variants
• RNA polymerase II
• Splicing speckles
• CRISPR mediated gene
• Clustered Regularly Interspaced Short Palindromic Repeats
• Loss of function