【资源】Ribozymes and siRNA protocols 2e (Humana Press)
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Table of Contents
1. Ribozyme- and siRNA-Mediated mRNA Degradaion: A General Introduction
2. Combination of Chemical and Enzymatic RNA Synthesis
3. Determination of Kinetic Parameters for Hammerhead and Hairpin Ribozymes
4. Real-Time Monitoring of RNA and DNA Reactions by Fluorescence Detection
5. Quantification of Ribozyme Target RNA Using Real-Time PCR
6. Analysis of Ribozyme Structure and Function by Nucleotide Analog Interference Mapping
7. Analysis of Global Conformational Transitions in RIbozymes
8. In Vivo Detection of Ribozyme Cleavage Products and RNA Structure by Use of Terminal Transferase-Dependent PCR
9. Identification of Efficient Cleavage Sites in Long-Target RNAs
10. Selection In Vitro of Allosteric Ribozymes
11. Regulation of Ribozyme Cleavage Activity by Oligonucleotides
12. Tetracycline-Regulated Expression of Hammerhead Ribozymes In Vivo
13. Ribozyme Expression Systems
14. Design and Expression of Chimeric U1/Ribozyme Transgenes
15. Design and Validation of Therapeutic Hammerhead Ribozymes for Autosomal Dominant Diseases
16. Helicase-Attached Novel Hybrid Ribozymes
17. Functional Gene Discovery Using Hybrid Ribozyme Libraries
18. Maxizyme Technology
19. Target-Site Selection for the 10-23 DNAzyme
20. In Vitro Selected RNA-Cleaving DNA Enzymes From Combinatorial Libraries
21. Nucleic Acid Sequence Analysis Using DNAzymes
22. Crystallization of the Hairpin Ribozyme: Illustrative Protocols
23. An Experimental Method for Selecting Effective Target Sites and Designing Hairpin Ribozymes
24. Design and Optimization of Sequence-Specific Hairpin Ribozymes
25. Design, Targeting, and Initial Screening of sTRSV-Derived Hairpin Ribozymes for Optimum Helix 1 Length and Catalytic Efficiency In Vitro
26. Optimization and Application of teh Group 1 Ribozyme Trans-Splicing Reaction
27. Repair of Myotonic Dystrophy Protein Kinase (DMPK) Transcripts by Trans-Splicing Ribozymes
28. General Design and Construction of RNase P Ribozymes for Gene-Targeting Application
29. In Vitro Selection of RNase P Ribozymes That Efficietnly Cleave a Target mRNA
30. In Vitro Selection of External Guide Sequences for Directing Human RNase P to Cleave a Target mRNA
31. RNase P-Mediated Inhibition of Viral Growth by Exogenous Administration of Short Oligonucleotide External Guide Sequence
32. RNase P Ribozyme As an Antiviral Agent Against Human Cytomegalovirus
33. Inhibition of Gene Expression by Nucleic Acid Enzymes in Rodent Models of Human Disease
34. Potential Design Rules an Enzymatic Synthesis of siRNAs
35. RNA Interference (RNAi) With RNase III-Prepared siRNAs
36. RNAi Expression Vectors in Mammalian Cells
37. Gene-Array Analysis of Glioma Cells After Treatment With Anti-PKC-a siRNA
38. RNAi in Living Mice
39. Construction and Transfection of PCR Products Expressing siRNAs or shRNAs in Mammalian Cells
40. Systemic Delivery of siRNAs
41. Adenovirus-Delivered siRNA
42. RNAi Expression Vectors in Plant Cells
43. Delivery Agents for Oligonucleotides
44. Selection of Peptides for Specific Delivery of Oligonucleotides Into Cancer Cells
45. Clinical Gene Therapy Research Utilizing Ribozymes: Application to the Treatment of HIV/AIDS
46. Critical Steps in the Implementation of Hematopoietic Progenitor-Cell Gene Therapy Using Ribozyme Vectors: A Laboratory Protocol
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Table of Contents
1. Ribozyme- and siRNA-Mediated mRNA Degradaion: A General Introduction
2. Combination of Chemical and Enzymatic RNA Synthesis
3. Determination of Kinetic Parameters for Hammerhead and Hairpin Ribozymes
4. Real-Time Monitoring of RNA and DNA Reactions by Fluorescence Detection
5. Quantification of Ribozyme Target RNA Using Real-Time PCR
6. Analysis of Ribozyme Structure and Function by Nucleotide Analog Interference Mapping
7. Analysis of Global Conformational Transitions in RIbozymes
8. In Vivo Detection of Ribozyme Cleavage Products and RNA Structure by Use of Terminal Transferase-Dependent PCR
9. Identification of Efficient Cleavage Sites in Long-Target RNAs
10. Selection In Vitro of Allosteric Ribozymes
11. Regulation of Ribozyme Cleavage Activity by Oligonucleotides
12. Tetracycline-Regulated Expression of Hammerhead Ribozymes In Vivo
13. Ribozyme Expression Systems
14. Design and Expression of Chimeric U1/Ribozyme Transgenes
15. Design and Validation of Therapeutic Hammerhead Ribozymes for Autosomal Dominant Diseases
16. Helicase-Attached Novel Hybrid Ribozymes
17. Functional Gene Discovery Using Hybrid Ribozyme Libraries
18. Maxizyme Technology
19. Target-Site Selection for the 10-23 DNAzyme
20. In Vitro Selected RNA-Cleaving DNA Enzymes From Combinatorial Libraries
21. Nucleic Acid Sequence Analysis Using DNAzymes
22. Crystallization of the Hairpin Ribozyme: Illustrative Protocols
23. An Experimental Method for Selecting Effective Target Sites and Designing Hairpin Ribozymes
24. Design and Optimization of Sequence-Specific Hairpin Ribozymes
25. Design, Targeting, and Initial Screening of sTRSV-Derived Hairpin Ribozymes for Optimum Helix 1 Length and Catalytic Efficiency In Vitro
26. Optimization and Application of teh Group 1 Ribozyme Trans-Splicing Reaction
27. Repair of Myotonic Dystrophy Protein Kinase (DMPK) Transcripts by Trans-Splicing Ribozymes
28. General Design and Construction of RNase P Ribozymes for Gene-Targeting Application
29. In Vitro Selection of RNase P Ribozymes That Efficietnly Cleave a Target mRNA
30. In Vitro Selection of External Guide Sequences for Directing Human RNase P to Cleave a Target mRNA
31. RNase P-Mediated Inhibition of Viral Growth by Exogenous Administration of Short Oligonucleotide External Guide Sequence
32. RNase P Ribozyme As an Antiviral Agent Against Human Cytomegalovirus
33. Inhibition of Gene Expression by Nucleic Acid Enzymes in Rodent Models of Human Disease
34. Potential Design Rules an Enzymatic Synthesis of siRNAs
35. RNA Interference (RNAi) With RNase III-Prepared siRNAs
36. RNAi Expression Vectors in Mammalian Cells
37. Gene-Array Analysis of Glioma Cells After Treatment With Anti-PKC-a siRNA
38. RNAi in Living Mice
39. Construction and Transfection of PCR Products Expressing siRNAs or shRNAs in Mammalian Cells
40. Systemic Delivery of siRNAs
41. Adenovirus-Delivered siRNA
42. RNAi Expression Vectors in Plant Cells
43. Delivery Agents for Oligonucleotides
44. Selection of Peptides for Specific Delivery of Oligonucleotides Into Cancer Cells
45. Clinical Gene Therapy Research Utilizing Ribozymes: Application to the Treatment of HIV/AIDS
46. Critical Steps in the Implementation of Hematopoietic Progenitor-Cell Gene Therapy Using Ribozyme Vectors: A Laboratory Protocol
