Term
| Any technique that uses living organisms to make or modify products, to improve plants or animals, or to develop microorganisms for specific uses |
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Definition
| Office of Technology Assessment 1991 Definition of Biotechnology |
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| Discovery of the fermentative feature of yeast by Louis Pasteur in 1857 |
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| Natural chemical compounds produced using microbial fermentation; Does NOT alter the genetic makeup of the organisms |
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| centered by passive techniques which lack efficacy and accuracy |
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| Traditional biotechnology |
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| uses gene manipulation to actively engineer organisms with high efficiency and accuracy |
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| Discovery of the double helix structure of DNA by James Watson and Francis Crick in 1953 |
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| Recominant DNA-- Genetic engineering |
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| Modern feature: manipulates genetic makeup |
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| produce biological compounds, traits or organisms that cannot be produced by natural processes or are not even naturally existent |
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| Not only for microorganisms but also for the large scale culture of mammalian cells |
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| Plant cell fusion creates new plants. Fusion of immunized spleen cells with myeloma cells forms hybridoma which produces: |
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| monoclonal antibodies (Cell fusion core technique) |
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| Basic research and clinical application |
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| manipulating the genetic constitution of organisms to make new proteins, express new traits, or correct genetic defects |
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| Recombinant DNA and gene transfer |
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| to generate genetically identical microorganisms or plants. Using nuclear transfer to generate genetically identical animals |
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| progenitor cells that are capable of self renewal and differentiation into many different cell lineages. Have potential for treatment of many malignant and non-malignant diseases |
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| Production of recombinant pharmaceuticals |
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| hormones, monoclonal antibodies, proteins, protein inhibitors; nucleic acid drugs |
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| disease diagnosis (applications of biotechnology) |
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Definition
| genomics, proteaomics, bioinformatics, -omics-based quick, accurate diagnosis |
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| applying recombinant DNA and transgenic techniques to modify and improve crop plants and farming animals |
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| Applications of Biotechnology: Healthcare and Pharmaceutics |
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| New drugs, new therapies, protective and therapeutic vaccines, disease diagnosis, personal medicine, agriculture, energy and environmental technology, food industry |
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| Nucleic Acid Modifications |
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Definition
| Phosphothioate DNA: PS; locked nucleic acid: LNA; 1'-0-methoxy-ethyl RNA: MOE; 2'-0-methyl RNA: OMe; 2'-Fluro of therapeutic nucleic acid analogs |
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| pharmacological significance of nucleic acid modifications |
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Definition
| increase stability of the nucleic acid drugs in vivo and improve their pharmacokinetics |
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| Classification of nucleic acid drugs |
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Definition
| Antisense oligonucleotides; siRNA; Aptamers; Ribozymes; CpG oligodeoxynucleotides (OCpG ODN) |
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Term
| short, synthetic, single-stranded DNA, RNA, or their analogues, designed to modulate gene expression by selective hybridization to complementary sequences in the target DNA, mRNA, or pre-mRNA |
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Definition
| Antisense oligonucleotides |
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| can form double stranded RNA molecule with sense mRNA to accelerate the degradation of mRNA or arrest translation by blocking ribosomal sites |
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| antisense oligonucleotide |
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| antisense oligos complementary to genomic DNA |
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Definition
| form triple helix structure to inhibit transcription |
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| Antisense RNA can enter transcription bubble |
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Definition
| then bind to their cognate sequence to inhibit transcription |
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| Antisense oligos as therapeutics |
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Definition
| can be synthesized easily, must be stable in vivo, must be able to enter the target cells, must not interact with other cellular target, should not interact in a non-sequence-specific manner with other macromolecules |
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Term
| small pieces of double stranded RNA with 3' overhangs at each end that can be used to interfere with the translation of proteins by binding to and promoting the degradation of messenger RNA at specific sequences |
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| used double stranded RNA to silence a specific gene in C. elegan. |
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| endogenous dsRNA that is encoded by microRNA gene |
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| member of ribonuclease III family |
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| RNA-induced silencing complex |
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| a member of Argonaute family, has endonuclease activity that cleave the mRNA into fragment |
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| Primary miRNA, shRNA or dsRNA |
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Definition
| binds to the protein Dicer, a component of RISC |
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| cleaves pri-miRNA, shRNA, dsRNA into 21-23 nt dsRNA fragment with 2 nt overhang at the 3' end |
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| One strand (guide strand) of the dsRNA |
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| The guide strand in the RISC associates with target mRNA |
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| by perfect or non-perfect Watson-Crick pairing |
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| is cleaved by AGO2 of RISC |
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| modulate protein function by directly binding to the target protein; much like monoclonal antibodies |
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| single stranded RNA or DNA that bind to molecular targets, ranging from small molecules to proteins, with high affinity and specificity due to their stable-three dimensional shapes |
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| many aptamers exists as hairpin-like monomers |
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Definition
| that bind targets via unpaired nucleotides |
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| Aptamers block or activate the target protein |
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Definition
| and regulate a specific signaling pathway via their specific and high-affinity binding to the target protein |
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Term
| Aptamers are evolved and identified in vitro using SELEX, |
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Definition
| can be reproducibly and economically synthesized in large scale for clinical applications |
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| Using chemical substitutions and other modifications (including l-RNA) |
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| aptamers elicit minimal immunogenicity relative to antibodies |
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Definition
| allows for improved transport and tissue penetration compared to antibodies |
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| aptamers are amenable to applications that require engineering |
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Definition
| such as the conjugation of aptamers to ribozymes and aptamers-siRNA chimeras |
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Term
| catalytics RNAs that function as enzymes and do not require proteins for catalysis |
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Definition
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| must contain a sequence that will bind to the target and a sequence that will fold into a structure with ribonuclease activity |
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| As therapeutic agents, the antisense region (substrate recognition domain) of the ribozyme |
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Definition
| can be designed to recognize specific target RNA sequence |
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| synthetic olidodeoxynucleotides containing unmethylated CpG motif that activate Toll-like receptor 9 (TLR( |
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Definition
| CpG Oligodeoxynucleotides (CpG ODNs) |
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| Induces exceptionally strong pDC IFN-alpha secretion and moderate expression of costimulatory molecules. Induces very little beta cell activation |
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Definition
| ODN: A-class (also called D-type) |
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| induces very strong beta cell proliferation and differentiation. Induces pOC expression of costimulatory molecules and modest iFN-alpha secretion |
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Definition
| ODN: B-class (also called K-type) |
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| induces strong B cell proliferation and differientiation; induces pDC IFN-alpha secretion and expression of costimulatory molecules |
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| is expressed in plasmacytoid Dendritic cells (pDC) and B cells; the activation induces pDC to produce large amount of IFN-alpha/beta which in turn induces T-helper 1 type immune response |
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| Therapeutic application of CpG ODNs |
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Definition
| Immunoprotective activity; Vaccine adjuvant; Prevention and treatment of allergy. |
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