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Antisense Technology

Antisense Technology. Delihas, N (2001) Antisense Nucleic Acids in Biotechnology. Encycloptedia of Life Sciences, Nature publishing. Nucleic Acid Drugs. Antisense oligonucleotides to block translation of a particular protein Small interfering RNAs micro RNAs RNA interference. Features.

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Antisense Technology

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  1. Antisense Technology Delihas, N (2001) Antisense Nucleic Acids in Biotechnology. Encycloptedia of Life Sciences, Nature publishing

  2. Nucleic Acid Drugs • Antisense oligonucleotides • to block translation of a particular protein • Small interfering RNAs • micro RNAs • RNA interference

  3. Features • Short synthetic oligonucleotides – DNA,RNA • Target • Cellular RNA • Specific block in transcription • RNA splicing/transport • translation • Viral RNA • Length to guarantee theoretical uniqueness • Human genome 3.0 x 109 bp 16 mer • Celluar RNA 1.5 x 107 bp 12 mer • Min. 12 mer Typically 18 mer Max. 24 mer • Modification to resist Dnase degradation • Inclusion of phosphorothioate groups

  4. Stability of RNA oligonucleotides • In 95% serum at 37℃

  5. Principal target sites • Transcription • DNA triple helix formation • Hybridization to DNA loops • Hybridization to nascent RNA • RNA processing • RNA splicing • RNA transport • Translation • Inhibition of initiation factor binding • Inhibition of ribosome assembly • Inhibition of movement

  6. Targeting • In theory : target AUG start codon • In practice : trial and error

  7. mRNA degradation • RNase H – ribonuclease that recognizes and cleaves mRNA in an RNA/DNA duplex

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