Ribozyme Protocols (eBook, PDF)

Redaktion: Turner, Philip C.

• Format: PDF

Produktbeschreibung

Ribozyme Protocols brings together a broad spectrum of essential and easily reproducible techniques currently used by leading researchers to manipulate RNA molecules and to inactivate gene expression in vivo. The methods collected here range from theoretical design to clinical application and include the synthesis and assay of ribozymes, as well as their use both in vitro and in vivo. Each method is presented by an expert with substantial hands-on experience and in far greater detail than is generally to be found in the research literature. Ribozyme Protocols will help all investigators perform meaningful and successful experiments using ribozymes in both clinical and basic research, as well as launch fresh application initiatives. It is certain to be found indispensable not only by those workers new to the field, but also by experienced researchers who wish to expand their range of expertise.

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Produktdetails

• Verlag: Humana Press
• Seitenzahl: 492
• Erscheinungstermin: 2. Februar 2008
• Englisch
• ISBN-13: 9781592595600
• Artikelnr.: 64700264

Inhaltsangabe

Ribozymes.- Computer-Aided Calculation of the Local Folding Potential of Target RNA and Its Use for Ribozyme Design.- Computational Approaches to the Identification of Ribozyme Target Sites.- Computer Analysis of the Conservation and Uniqueness of Ribozyme-Targeted HIV Sequences.- Selection of Accessible Sites for Ribozymes on Large RNA Transcripts.- Selection of Efficient Ribozyme Cleavage Sites in Target RNAs.- Chemical Synthesis, Analysis, and Purification of Ribozymes.- A Practical Method for the Production of RNA and Ribozymes.- Preparation of Templates for Production of Ribozymes and Substrates.- Enzymatic Synthesis and Characterization of Unmodified Ribozymes and Substrates.- T7 Transcript Length Determination Using Enzymatic RNA Sequencing.- Chemical and Enzymatic Approaches to Construct Modified RNAs.- Applications of Modified Transcripts.- Cloning Strategies for Catalytic Antisense RNAs.- PCR-Based Construction of Long Hammerhead Ribozymes.- Design and Production of Asymmetric Hammerhead Ribozymes.- Minimized Hammerhead Ribozymes.- Design of Hairpin Ribozymes for In Vitro and Cellular Applications.- Design of the Hairpin Ribozyme for Targeting Specific RNA Sequences.- Design and Preparation of Sequence-Specific RNase P Ribozymes.- Theoretical Considerations in Measuring Reaction Parameters.- Experimental Approaches for Measuring Reaction Parameters.- Determination of Catalytic Parameters for Hairpin Ribozymes.- Characterizing Ribozyme Cleavage Reactions.- Defining Optimum Reaction Conditions for Hammerhead Ribozymes.- Using Fluorescence Resonance Energy Transfer to Investigate Hammerhead Ribozyme Kinetics.- Design of Hybridizing Arms in Hammerhead Ribozymes.- Optimization of Hammerhead Flanking Sequences Using Oligonucleotide Facilitators.- Enhancement of Ribozyme Function by RNA Binding Proteins.- Selection of Fast-Hybridizing Complementary RNA Species In Vitro.- In Vitro Selection of Hairpin Ribozymes.- The Detection of Hammerhead Ribozyme Cleavage by RT-PCR Methods.- Detection of Ribozyme Cleavage Products Using Reverse Ligation-Mediated PCR (RL-PCR).- Quantitation of Ribozyme Target Abundance by QCPCR.- Trans-Splicing Reactions by Ribozymes.- Ligation of RNA Molecules by the Hairpin Ribozyme.- Mutagenesis and Modeling of the Hairpin Ribozyme Family.- Preparation of Homogeneous Ribozyme RNA for Crystallization.- Establishing Suitability of RNA Preparations for Crystallization.- A Sparse Matrix Approach to Crystallizing Ribozymes and RNA Motifs.- Crystallographic Analyses of Chemically Synthesized Modified Hammerhead RNA Sequences as a General Approach Toward Understanding Ribozyme Structure and Function.- tRNA Delivery Systems for Ribozymes.- Expressing Ribozymes in Plants.- Using Microinjection of Xenopus Oocytes to Express and Optimize Ribozymes In Vivo.- Exogenous Cellular Delivery of Ribozymes and Ribozyme Encoding DNAs.- Optimization of Lipid-Mediated Ribozyme Delivery to Cells in Culture.- Retroviral Delivery and Anti-HIV Testing of Hammerhead Ribozymes.- Hairpin Ribozyme Gene Therapy for AIDS.- Clinical Aspects of Ribozymes as Therapeutics in Gene Therapy.