Gene Targeting Protocols

ISBN-13: 9780896033603 / Angielski / Twarda / 1999 / 233 str.

Eric B. Kmiec; Dieter C. Gruenert

Opis

The potential now exists in many experimental systems to transfer a cloned, modified gene back into the genome of the host organism. In the ideal situation, the cloned gene is returned to its homologous location in the genome and becomes inserted at the target locus. This process is a controlled means for the repair of DNA damage and ensures accurate chromosome disjunction during meiosis. The paradigm for thinking about the mechanism of this p- cess has emerged primarily from two sources: (1) The principles of reaction mechanics have come from detailed biochemical analyses of the RecA protein purified from Escherichia coli; and (2) the principles of information transfer have been derived from genetic studies carried out in bacteriophage and fungi. A compelling picture of the process of homologous pairing and DNA strand exchange has been influential in directing investigators interested in gene t- geting experiments. The ability to find and pair homologous DNA molecules enables ac- rate gene targeting and is the central phenomenon underlying genetic recombi- tion. Biochemically, the overall process can be thought of as a series of steps in a reaction pathway whereby DNA molecules are brought into homologous register, the four-stranded Holliday structure intermediate is formed, hete- duplex DNA is extended, and DNA strands are exchanged. Not much is known about the biochemical pathway leading to homologous recombination in euka- otes.

Recenzje

"The book also has several techniques for using oligonucleotides in gene targeting, a methodology that may well revolutionize the entire field." - Cellular and Molecular Biology

Spis treści

Nucleic Acid Transfer Using Cationic Lipids, Natasha J. Caplen. Optimizing Polyethylenimine-Based Gene Transfer into Mammalian Brain for Analysis of Promoter Regulation and Protein Function, Barbara A. Demeneix, Mohamed Ghorbel, and Daniel Goula. Gene Transfer and Drug Delivery by Electronic Pulse Delivery: A Nonviral Delivery Approach, Xi Zhao. Strategies for Improving the Frequency and Assessment of Homologous Recombination, Nancy Smyth Templeton. Effective Gene Transfer Using Viral Vectors Based on SV40, David S. Strayer. Rapid Generation of Isogenic Mammalian Cell Lines Expressing Recombinant Transgenes by Use of Cre Recombinase, Bruce D. Bethke and Brian Sauer. Site-Directed Alteration of DNA by Small-Fragment Homologous Replacement, Kaarin K. Goncz and Dieter C. Gruenert. Mutation Correction by Homologous Recombination with an Adenovirus Vector, Ayumi Fujita-Kusano, Yasuhiro Naito, Izumu Saito, and Ichizo Kobayashi. Site-Specific Targeting of DNA Plasmids to Chromosome 19 Using AAV Cis and Trans Sequences, Samuel M. Young Jr., Weidong Xiao, and Richard Jude Samulski. Adeno-Associated Virus Based Gene Therapy in Skeletal Muscle, Richard J. Bartlett and Jesica M. McCue. Rapid Establishment of Myeloma Cell Lines Expressing Fab(Tac)-Protamine, a Targetable Protein Vector, Directed Against High-Affinity a-Chain of Human Interleukin-2 Receptor, Sun U. Song and Wayne A. Marasco. EBV-Derived Episomes to Probe Chromatin Structure and Gene Expression in Human Cells, Juliette Fivaz, M. Chiara Bassi, Stéphane Pinaud, Larry Richman, Melanie Price, and Jovan Mirkovitch. Triplex-Directed Site-Specific Genome Modification, Karen M. Vasquez and John H. Wilson. Use of Quantitative Ligation-Mediated Polymerase Chain Reaction to Detect Gene Targeting by Alkylating Oligodeoxynucleotides, Howard B. Gamper, Irina Afonina, Evgeniy Belousov, Michael W. Reed, and Mikhail A. Podyminogin. Gene Targeting in Plants via Site-Directed Mutagenesis, Peter B. Kipp, Joyce M. Van Eck, Peter R.Beetham, and Gregory D. May. Antisense Oligonucleotides as Modulators of Pre-mRNA Splicing, Halina Sierakowska, Sudhir Agrawal, and Ryszard Kole. Index.

Blurb

In Gene Targeting Protocols, a panel of innovative investigators present in readily reproducible detail the latest techniques for gene replacement, gene knockout, and gene repair in both animals and plants. These expert researchers review the best gene transfer vehicles (liposomes, PEI, and EPD), discuss the parameters that affect vector uptake, and detail successful methods for gene targeting (Cre-ox, and adeno and adeno-associated viruses). The methods include two fundamentally different approaches to gene targeting- one using a fusion protein to deliver a vector to a virus for possible therapeutic effect, and one entailing an episomal-based Epstein-Barr vector that can modulate the chromatin assembly process, and thus overcome a serious barrier to therapeutic gene targeting. The book also has several techniques for using oligonucleotides in gene targeting, a methodology that may revolutionize the entire field.
Innovative and wide ranging, Gene Targeting Protocols offers today's researchers robust and proven gene-targeting techniques that are essential to understanding and regulating biological processes at the genetic level.