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A. THE GLOBAL SCENE
RNA Interference and RNA Silencing
•    RNA Interference (RNAi)- is a highly
    evolutionally conserved process of post-
    transcriptional gene silencing (PTGS) by
    which double stranded RNA (dsRNA),
    when introduced into a cell, causes
    sequence-specific degradation of
    homogolous mRNA sequences.
• 1998- it was introduced by Andrew Fire
  and Craig Mello in the nematode worm
      Caenorhabditis elegans and later found
  in a wide variety of organisms, including
      mamals.

•      RNAi is believed to be universal as it
    has been observed in all eukaryotes, from
    yeast to     mamals.
The most interesting aspects of RNAi are
   the following:
• dsRNA, rather than single-stranded
   antisense RNA, is the interfering agent.
• It is highly specific
• It is remarkably potent
• the interfering activity can cause
   interference in cells and tissues far
   removed from the site of introduction.
Application of RNAi

1. Determining Gene Function
2. Pathway Analysis
3. Identify and Validate drug Targets
4. Study Gene Redundancy
5. Functional Screening
RNA Timeline
Timelin         Who                        What                  Organism
   e

          Guo S, and   First noticed that sense RNA was
1995      Kemphues KJ. as effective as antisense                 C. elegans
                            RNA for Suppressing Gene
                            expression in worm
                            First described RNAi Phenomenon
1998      Fire et al.       in C. elegans by injecting dsRNA
                            into C. elegans which led to an      C. elegans
                            efficient sequence- specific
                            silencing and coined the term
                            “RNA Interference”.

                            Reported processing of long dsRNA
2000        Zamore et al.   by Rnase II (Dicer) into shorter     Drosophila
                            fragments of 21-23-nt intervals in
                            Drosophila extracts
Cloned Dicer, the Rnase III enzyme that is
2001   Bernstein et al.   evolutionarily conserved and contains        C. elegans
                          helicase and PAZ domains, as well as two
                          dsRNA- Binding domains.

2001   Tuschl T and                                                    Mammals
       colleagues         First described RNAi in Mammalian cells.

2003   Paddison et al.    Short hairpin RNAs (shRNAs) induce           Mammals
       Sui et al.         sequence- specific silencing in mamalian
                          cells.
       Paul et al.

2003                      First reported that siRNAs can be used       Mammals
       Song et al.        therapeutically in whole animals.

2004   Kawasaki and       First observed that siRNA silences gene at   Human
       Taira              transcriptional level possibly through
       Morris et al.      directing de novo DNA methylation.

       Andrew Fire        Won Noble Prize in Physiology or Medicine    Human
2006   and                for discovering RNAi mechanism.
       Craig Mello
2006   Li et al           First reported that small dsRNA induces      Human
                          gene expresion activation, a phenomenon
                          termed as RNAa.
DNA Microarray Technology?
•    DNA Microarray Technology- have
    been developed in the 1990's as a
    method of rapidly analyzing the
    expression of thousands of genes all at
    the same time by allowing the RNA
    products of genes to be monitored at
    once.
How do DNA microarrays work?
•     DNA microarray work by providing
  fixed single strands of Dna (gene
  sequences) to which labeled cDNA
  fragments can bind. Thousands of
  different genes are immobilized onto ma
  small, inert, solid support (called a chip),
  which is usually a glass microscope slide,
  but can also be a silicon chip or a nylon
  membrane.
Application of DNA microarrays work?

•   Correlating Gene Expression with
    metabolic Changes
•   Characterizing nCell Types
•   Cancer Research
•   Toxicology
FBI CODIS (Combined DNA Index
    System) for Human Identification
•    In 1997 the US FBI (Federal Bureau of
    Investigation) Selected thirteen STR
    (Short Tandem Repeat) loci in human
    chromosomes foe use as standards in
    human identification and genome
    signature comparison.
Why use STRs (Short Tandem Repeats)?
•   Short Tandem Repeats (STRs) offer
  several advantages over RFLPs as tools
  for human identification.
• STRs exhibit high heterozygosity
• STRs exhibit a regular repeat pattern
• STRs are easily amplified
IV. Nanotechnology
•    Nanotechnology- refers to the design,
  characterization, production and
  application of structures, devices and
  systems by controlling shape and size at
  nanometer scale.
• Nano- is derived from the greek word
  “dwarf”.
Bioinformatics: Shaping the Future of
                 Biotechnology

•      Blast- is an algorithm that stands for
      Basic Local Alignment Search Tool and
      was developed by Altschul, Gish, Miller,
      Myers, and Lipman in 1990.
•     It is used to search for related
      sequences available in the database
      and for comparing two or more
      sequences for similarities.
Application in Biotechnology
                    Research
•   Agriculture
•   Aquaculture
•   Human Health
•   Environment
•   Wildlife Conservation
•   Industry
•   Forensics

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The global scene

  • 2. RNA Interference and RNA Silencing • RNA Interference (RNAi)- is a highly evolutionally conserved process of post- transcriptional gene silencing (PTGS) by which double stranded RNA (dsRNA), when introduced into a cell, causes sequence-specific degradation of homogolous mRNA sequences.
  • 3. • 1998- it was introduced by Andrew Fire and Craig Mello in the nematode worm Caenorhabditis elegans and later found in a wide variety of organisms, including mamals. • RNAi is believed to be universal as it has been observed in all eukaryotes, from yeast to mamals.
  • 4. The most interesting aspects of RNAi are the following: • dsRNA, rather than single-stranded antisense RNA, is the interfering agent. • It is highly specific • It is remarkably potent • the interfering activity can cause interference in cells and tissues far removed from the site of introduction.
  • 5. Application of RNAi 1. Determining Gene Function 2. Pathway Analysis 3. Identify and Validate drug Targets 4. Study Gene Redundancy 5. Functional Screening
  • 7. Timelin Who What Organism e Guo S, and First noticed that sense RNA was 1995 Kemphues KJ. as effective as antisense C. elegans RNA for Suppressing Gene expression in worm First described RNAi Phenomenon 1998 Fire et al. in C. elegans by injecting dsRNA into C. elegans which led to an C. elegans efficient sequence- specific silencing and coined the term “RNA Interference”. Reported processing of long dsRNA 2000 Zamore et al. by Rnase II (Dicer) into shorter Drosophila fragments of 21-23-nt intervals in Drosophila extracts
  • 8. Cloned Dicer, the Rnase III enzyme that is 2001 Bernstein et al. evolutionarily conserved and contains C. elegans helicase and PAZ domains, as well as two dsRNA- Binding domains. 2001 Tuschl T and Mammals colleagues First described RNAi in Mammalian cells. 2003 Paddison et al. Short hairpin RNAs (shRNAs) induce Mammals Sui et al. sequence- specific silencing in mamalian cells. Paul et al. 2003 First reported that siRNAs can be used Mammals Song et al. therapeutically in whole animals. 2004 Kawasaki and First observed that siRNA silences gene at Human Taira transcriptional level possibly through Morris et al. directing de novo DNA methylation. Andrew Fire Won Noble Prize in Physiology or Medicine Human 2006 and for discovering RNAi mechanism. Craig Mello 2006 Li et al First reported that small dsRNA induces Human gene expresion activation, a phenomenon termed as RNAa.
  • 9. DNA Microarray Technology? • DNA Microarray Technology- have been developed in the 1990's as a method of rapidly analyzing the expression of thousands of genes all at the same time by allowing the RNA products of genes to be monitored at once.
  • 10. How do DNA microarrays work? • DNA microarray work by providing fixed single strands of Dna (gene sequences) to which labeled cDNA fragments can bind. Thousands of different genes are immobilized onto ma small, inert, solid support (called a chip), which is usually a glass microscope slide, but can also be a silicon chip or a nylon membrane.
  • 11. Application of DNA microarrays work? • Correlating Gene Expression with metabolic Changes • Characterizing nCell Types • Cancer Research • Toxicology
  • 12. FBI CODIS (Combined DNA Index System) for Human Identification • In 1997 the US FBI (Federal Bureau of Investigation) Selected thirteen STR (Short Tandem Repeat) loci in human chromosomes foe use as standards in human identification and genome signature comparison.
  • 13. Why use STRs (Short Tandem Repeats)? • Short Tandem Repeats (STRs) offer several advantages over RFLPs as tools for human identification. • STRs exhibit high heterozygosity • STRs exhibit a regular repeat pattern • STRs are easily amplified
  • 14. IV. Nanotechnology • Nanotechnology- refers to the design, characterization, production and application of structures, devices and systems by controlling shape and size at nanometer scale. • Nano- is derived from the greek word “dwarf”.
  • 15. Bioinformatics: Shaping the Future of Biotechnology • Blast- is an algorithm that stands for Basic Local Alignment Search Tool and was developed by Altschul, Gish, Miller, Myers, and Lipman in 1990. • It is used to search for related sequences available in the database and for comparing two or more sequences for similarities.
  • 16. Application in Biotechnology Research • Agriculture • Aquaculture • Human Health • Environment • Wildlife Conservation • Industry • Forensics