21ntpassenger5′→3′-cloudfrontnet.ppt

* * * * Dr. Prashant Kumar Asst. Professor Amity Institute of Virology Immunology Amity University Uttar Pradesh Nucleic acid based multi-target approach for effective silencing of influenza A virus INFLUENZA One of the major Respiratory Virus Causative agent of one of the most common disease i.e. common cold/ fever Most easily transmissible disease- through aerosol Strains keep on changing every season Cause of over 2,00,000 deaths every year INFLUENZA Types of Influenza Virus A B C H1N1 H3N2 (In Humans) (In Humans) Non Virulent H5N1 (In Birds) HOST RANGE: Humans Swine Birds (Wild aquatic birds-Reservoir) Horses Whales Seals Influenza Virus-Prone to Mutation Nature Reviews/ Microbiology Control Options for Influenza: Vaccines: Seasonal vaccine (Trivalent/ Quadrivalent) Drugs: Oseltamivir (Tamiflu) Zanamivir (Relenza) Peramivir (Rapivab) Amantadine Rimantadine Nucleic Acids based therapeutics Therapeutic Tools siRNA/shRNA Ribozymes DNAzymes Hasnoot et al 2007 5’ NNNNNNNUX NNNNNNN YYYYYYYA YYYYYYY A CUG A A G AGU CG AU GC GC A G GU 3’ 5’ 3’ catalytic motif target mRNA Rz cleavage site Designing of Hammer-head Ribozyme Designing of siRNA Rules for selecting siRNA targets on mRNA sequences: Targets should be located 50-100 nt downstream of the start codon (ATG). Search for sequence motif AA(N19)TT or NA(N21), or NAR(N17)YNN, where N is any nucleotide, R is purine (A, G) and Y is pyrimidine (C, U). Target sequences should have a G+C content between 35-60%. Avoid stretches of 4 or more nucleotide repeats. Avoid 5URT and 3UTR, although siRNAs targeting UTRs have been shown to successfully induce gene silencing. Avoid sequences that share a certain degree of homology with other related or unrelated genes. Segment Size (nt) Polypeptide(s)