Author: Jason Robert Epstein
Publisher:
ISBN:
Category : HIV (Viruses)
Languages : en
Pages : 136
Book Description
Novel Methodology for the Linking of Oligonucleotide-peptide Conjugates
Author: Jason Robert Epstein
Publisher:
ISBN:
Category : HIV (Viruses)
Languages : en
Pages : 136
Book Description
Publisher:
ISBN:
Category : HIV (Viruses)
Languages : en
Pages : 136
Book Description
Development of a Novel Methodology for the Synthesis of Oligonucleotide-peptide Conjugates
Author: Simone Zaramella
Publisher:
ISBN: 9789173499194
Category :
Languages : en
Pages : 60
Book Description
Publisher:
ISBN: 9789173499194
Category :
Languages : en
Pages : 60
Book Description
Oligonucleotide Synthesis
Author: Piet Herdewijn
Publisher: Springer Science & Business Media
ISBN: 1592598234
Category : Medical
Languages : en
Pages : 436
Book Description
A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression.
Publisher: Springer Science & Business Media
ISBN: 1592598234
Category : Medical
Languages : en
Pages : 436
Book Description
A collection of powerful new techniques for oligonucleotide synthesis and for the use of modified oligonucleotides in biotechnology. Among the protocol highlights are a novel two-step process that yields a high purity, less costly, DNA, the synthesis of phosphorothioates using new sulfur transfer agents, the synthesis of LNA, peptide conjugation methods to improve cellular delivery and cell-specific targeting, and triple helix formation. The applications include using molecular beacons to monitor the PCR amplification process, nuclease footprinting to study the sequence-selective binding of small molecules of DNA, nucleic acid libraries, and the use of small interference RNA (siRNA) as an inhibitor of gene expression.
Bioconjugate Techniques
Author: Greg T. Hermanson
Publisher: Elsevier
ISBN: 0080527906
Category : Medical
Languages : en
Pages : 813
Book Description
Bioconjugate Techniques is the essential guide to the modification and crosslinking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or crosslinking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. Armed with this information and the abundant protocols provided, readers will form unique complexes that can be used for detecting, quantifying, and targeting important analytes. This book helps readers make: high activity antibody-enzymes conjugates, immunotoxins, immunogen complexes, liposome conjugates; as well as biotinylated molecules, avidin or streptavidin conjugates, colloidal gold labeled proteins, PEG or dextran complexes, labeled oligonucleotide probes, and fluorescently tagged or radiolabeled molecules. This book is the first to thoroughly capture the entire field of bioconjugate chemistry in a single volume Serves as a practical guide to modification and cross-linking technology for research, diagnostics, and therapeutics Provides useful, detailed, easy-to-follow, step-by-step protocols Contains easy-to-read, and easy-to-understand key concepts for making bioconjugates of all types Efficiently covers the chemistry of bioconjugation, the major reagents available for modification and cross-linking, and the application of these reagents to the synthesis of highly active conjugates Cites over more than references keyed to concepts covered in the book Uses more than 600 figures to illustrate bioconjugate reagents, their reactions, and applications Suggests sources for all key reagents
Publisher: Elsevier
ISBN: 0080527906
Category : Medical
Languages : en
Pages : 813
Book Description
Bioconjugate Techniques is the essential guide to the modification and crosslinking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or crosslinking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. Armed with this information and the abundant protocols provided, readers will form unique complexes that can be used for detecting, quantifying, and targeting important analytes. This book helps readers make: high activity antibody-enzymes conjugates, immunotoxins, immunogen complexes, liposome conjugates; as well as biotinylated molecules, avidin or streptavidin conjugates, colloidal gold labeled proteins, PEG or dextran complexes, labeled oligonucleotide probes, and fluorescently tagged or radiolabeled molecules. This book is the first to thoroughly capture the entire field of bioconjugate chemistry in a single volume Serves as a practical guide to modification and cross-linking technology for research, diagnostics, and therapeutics Provides useful, detailed, easy-to-follow, step-by-step protocols Contains easy-to-read, and easy-to-understand key concepts for making bioconjugates of all types Efficiently covers the chemistry of bioconjugation, the major reagents available for modification and cross-linking, and the application of these reagents to the synthesis of highly active conjugates Cites over more than references keyed to concepts covered in the book Uses more than 600 figures to illustrate bioconjugate reagents, their reactions, and applications Suggests sources for all key reagents
Novel Approaches to Oligonucleotide Conjugation to a Cell Penetrating (Tat) Peptide
Author: Victoria Steven
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
Book Description
Modifed oligonucleotides are routinely employed as bioanalytical probes for use in diagnostics; however, a major limiting factor in oligonucleotide-based diagnostics is poor cellular uptake. This can be overcome by covalent attachment of a cell penetrating peptide. Diels-Alder cycloaddition is an attractive methodology for oligonucleotide peptide conjugation; the reaction is fast and chemoselective and the reaction rate is greatly enhanced in aqueous media. An oligonucleotide sequence has been derivatised with a series of dienes at the 5'- terminus, through chemical synthesis of a series of unique dienyl modified phosphoramidites. Investigation into the effect of diene type on the efficiency of conjugation to a maleimido derivatised Tat peptide derivative, via the Diels-Alder cycloaddition, has been performed. The optimal diene for biomolecule conjugation was found to be cyclohexadiene in the conjugation of oligonucleotides to a cell penetrating peptide via Diels-Alder cycloaddition for the first time. An oligonucleotide sequence has also been derivatised with cyclohexadiene internally; Diels-Alder cycloadditions of these oligonucleotides to a maleimido derivatised Tat peptide derivative were also successful. However, oligonucleotide conjugation to Tat peptide via Diels-Alder cycloaddition of fluorescently labelled oligonucleotides for visualisation in cell studies has not been as successful as for unlabelled oligonucleotides. Generation of a biocatalytic DNA sequence for the acceleration of Diels-Alder cycloadditions has been attempted. Through a SELEX-type process, a DNA sequence has been isolated for experimental determination of its potential as a biological catalyst for Diels-Alder cycloadditions. Oligonucleotide conjugation to Tat peptide via gold nanoparticles has been achieved. Chemical synthesis of a dithiolated, long-chain PEGylated ligand allowed covalent attachment of Tat peptide to gold nanoparticles. Bifunctionalisation of gold nanoparticles with oligonucleotides and this ligand, followed by covalent attachment of Tat peptide generated the desired conjugate. The conjugates have been shown to hybridise successfully with the complementary oligonucleotide sequence, thereby displaying potential for their use as bioanalytical probes. Methods for quantification of both oligonucleotides and Tat peptide conjugated to gold nanoparticles, by enzyme hydrolysis, have been developed.
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
Book Description
Modifed oligonucleotides are routinely employed as bioanalytical probes for use in diagnostics; however, a major limiting factor in oligonucleotide-based diagnostics is poor cellular uptake. This can be overcome by covalent attachment of a cell penetrating peptide. Diels-Alder cycloaddition is an attractive methodology for oligonucleotide peptide conjugation; the reaction is fast and chemoselective and the reaction rate is greatly enhanced in aqueous media. An oligonucleotide sequence has been derivatised with a series of dienes at the 5'- terminus, through chemical synthesis of a series of unique dienyl modified phosphoramidites. Investigation into the effect of diene type on the efficiency of conjugation to a maleimido derivatised Tat peptide derivative, via the Diels-Alder cycloaddition, has been performed. The optimal diene for biomolecule conjugation was found to be cyclohexadiene in the conjugation of oligonucleotides to a cell penetrating peptide via Diels-Alder cycloaddition for the first time. An oligonucleotide sequence has also been derivatised with cyclohexadiene internally; Diels-Alder cycloadditions of these oligonucleotides to a maleimido derivatised Tat peptide derivative were also successful. However, oligonucleotide conjugation to Tat peptide via Diels-Alder cycloaddition of fluorescently labelled oligonucleotides for visualisation in cell studies has not been as successful as for unlabelled oligonucleotides. Generation of a biocatalytic DNA sequence for the acceleration of Diels-Alder cycloadditions has been attempted. Through a SELEX-type process, a DNA sequence has been isolated for experimental determination of its potential as a biological catalyst for Diels-Alder cycloadditions. Oligonucleotide conjugation to Tat peptide via gold nanoparticles has been achieved. Chemical synthesis of a dithiolated, long-chain PEGylated ligand allowed covalent attachment of Tat peptide to gold nanoparticles. Bifunctionalisation of gold nanoparticles with oligonucleotides and this ligand, followed by covalent attachment of Tat peptide generated the desired conjugate. The conjugates have been shown to hybridise successfully with the complementary oligonucleotide sequence, thereby displaying potential for their use as bioanalytical probes. Methods for quantification of both oligonucleotides and Tat peptide conjugated to gold nanoparticles, by enzyme hydrolysis, have been developed.
Therapeutic Oligonucleotides
Author: Jens Kurreck
Publisher: Royal Society of Chemistry
ISBN: 0854041168
Category : Medical
Languages : en
Pages : 362
Book Description
This book provides a compelling overall update on current status of RNA interference
Publisher: Royal Society of Chemistry
ISBN: 0854041168
Category : Medical
Languages : en
Pages : 362
Book Description
This book provides a compelling overall update on current status of RNA interference
Peptide Conjugation to Enhance Oligonucleotide Delivery
Author: Justin Mahoney Wolfe
Publisher:
ISBN:
Category :
Languages : en
Pages : 419
Book Description
The intracellular delivery of functional macromolecules remains an outstanding challenge in biomedicine. While small molecules can diffuse through the plasma membrane, many large therapeutic molecules are not internalized to an appreciable extent. One strategy to improve cell uptake involves linking the molecule of interest to a cell-penetrating peptide (CPP). CPPs are widely employed to enhance macromolecule delivery, with hundreds of different peptides and modifications reported to improve cellular uptake. In this thesis, CPPs were systematically investigated and chemically altered to facilitate the delivery of antisense oligonucleotides. To accurately compare the existing CPPs, 64 CPP sequences were synthesized, conjugated to oligonucleotides, and assayed for delivery. These CPPs showed a range of effectiveness, with some CPPs hindering the delivery of oligonucleotide cargo and others leading to a 10-fold increase in oligonucleotide activity. To help identify which CPPs might be valuable for oligonucleotide delivery specifically, a computational model was developed to predict, de novo, whether or not a CPP will be effective. When experimentally validated, this model successfully predicted which sequences would improve oligonucleotide delivery greater than 3-fold. Multiple strategies were employed to improve CPP effectiveness. First, arginine-rich CPPs were chemically modified with perfluoroarenes. Cyclic and bicyclic CPPs were synthesized by linking multiple cysteine residues together with a perfluoroarene. After oligonucleotide conjugation, these peptides led to a 14-fold increase in delivery. Second, two different CPPs were combined into one long chimeric sequence. The CPP chimeras were highly active, leading to a 20-fold increase in oligonucleotide delivery. Third, the idea of combining multiple CPPs led to the development of a method for the rapid synthesis combinatorial peptide conjugates. Using the judicious choice of bioconjugation chemistry, highly-active modular constructs were synthesized that contain three peptides linked to one oligonucleotide. In addition to CPPs for oligonucleotide delivery, one section of this thesis employed perfluoroaryl macrocyclic peptides to address the challenge of peptide delivery across the blood-brain barrier. An additional section developed a new peptide conjugation strategy that uses palladium-peptide oxidative addition complexes as solid, storable, and water-soluble reagents for bioconjugation.
Publisher:
ISBN:
Category :
Languages : en
Pages : 419
Book Description
The intracellular delivery of functional macromolecules remains an outstanding challenge in biomedicine. While small molecules can diffuse through the plasma membrane, many large therapeutic molecules are not internalized to an appreciable extent. One strategy to improve cell uptake involves linking the molecule of interest to a cell-penetrating peptide (CPP). CPPs are widely employed to enhance macromolecule delivery, with hundreds of different peptides and modifications reported to improve cellular uptake. In this thesis, CPPs were systematically investigated and chemically altered to facilitate the delivery of antisense oligonucleotides. To accurately compare the existing CPPs, 64 CPP sequences were synthesized, conjugated to oligonucleotides, and assayed for delivery. These CPPs showed a range of effectiveness, with some CPPs hindering the delivery of oligonucleotide cargo and others leading to a 10-fold increase in oligonucleotide activity. To help identify which CPPs might be valuable for oligonucleotide delivery specifically, a computational model was developed to predict, de novo, whether or not a CPP will be effective. When experimentally validated, this model successfully predicted which sequences would improve oligonucleotide delivery greater than 3-fold. Multiple strategies were employed to improve CPP effectiveness. First, arginine-rich CPPs were chemically modified with perfluoroarenes. Cyclic and bicyclic CPPs were synthesized by linking multiple cysteine residues together with a perfluoroarene. After oligonucleotide conjugation, these peptides led to a 14-fold increase in delivery. Second, two different CPPs were combined into one long chimeric sequence. The CPP chimeras were highly active, leading to a 20-fold increase in oligonucleotide delivery. Third, the idea of combining multiple CPPs led to the development of a method for the rapid synthesis combinatorial peptide conjugates. Using the judicious choice of bioconjugation chemistry, highly-active modular constructs were synthesized that contain three peptides linked to one oligonucleotide. In addition to CPPs for oligonucleotide delivery, one section of this thesis employed perfluoroaryl macrocyclic peptides to address the challenge of peptide delivery across the blood-brain barrier. An additional section developed a new peptide conjugation strategy that uses palladium-peptide oxidative addition complexes as solid, storable, and water-soluble reagents for bioconjugation.
Protocols for Oligonucleotide Conjugates
Author: Sudhir Agrawal
Publisher: Humana Press
ISBN: 9780896032521
Category : Science
Languages : en
Pages : 377
Book Description
Publisher: Humana Press
ISBN: 9780896032521
Category : Science
Languages : en
Pages : 377
Book Description
Self-assembly of Novel Peptide-oligonucleotide Conjugates
Author: Agata Chotera
Publisher:
ISBN:
Category :
Languages : en
Pages : 91
Book Description
"In this study, we were able to introduce for the first time a self-organizing system based on amphiphilic peptides and DNA-pep chimeras and describe a variety of supramolecular structures emerging in their mixtures... The presented research is of great importance in the field of bio-inspired supramolecular materials, as well as the origins of life." -- from abstract.
Publisher:
ISBN:
Category :
Languages : en
Pages : 91
Book Description
"In this study, we were able to introduce for the first time a self-organizing system based on amphiphilic peptides and DNA-pep chimeras and describe a variety of supramolecular structures emerging in their mixtures... The presented research is of great importance in the field of bio-inspired supramolecular materials, as well as the origins of life." -- from abstract.
The Synthesis of Oligonucleotide Peptide Conjugates Using Diels-Alder Chemistry
Author: Leann Tait
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
Book Description
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
Book Description