Ulteriori informazioni
This detailed volume explores the notable progress in the field of zinc finger proteins (ZFP) study through widely used methods and protocols involving their biological functions and applications. Beginning with a section on the basic biology of ZFPs and design and applications of custom ZFPs, the book continues by covering methods for the evaluation and prevention of ZFN-mediated cytotoxicity as well as a collection of the representative methods of ZFN delivery. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Zinc Finger Proteins: Methods and Protocols aims to prompt both the understanding of ZFP biology and the development of next-generation ZFP therapeutics.
Sommario
Part 1. Basic Biology and Engineering of ZFPs.- PRDM14, a zinc finger protein, regulates cancer stemness.- Computational methods for analysis of the DNA-binding preferences of Cys2His2 zinc finger proteins.- Design and application of mA-specific Zinc finger proteins for the readout of DNA methylation.- Enhanced manipulation of human mitochondrial DNA heteroplasmy in vitro using tunable mtZFN technology.- Engineering RNA-binding proteins by modular assembly of RanBP2-type Zinc Fingers.- Design of a system for monitoring ubiquitination activities of E2 enzymes using engineered RING finger proteins.- Directed evolution of targeted recombinases for genome engineering.- Optical recording of cellular zinc dynamics with zinc finger based biosensors.- Delivery of superoxide dismutase using Cys2-His2 zinc-finger proteins.- Genome editing of MSCs as a platform for cell therapy.- Part 2. Methods for the Evaluation and Prevention of ZFN Cytotoxicity.- Integrated multimodal evaluation of genotoxicity in ZFN-modified primary human cells.- Selection and characterization of DNA aptamers against FokI nuclease domain.- An improved genome engineering method using surrogate reporter-coupled suicidal ZFNs.- Part 3. Delivery Methods of ZFPs.- Non-transgenic approach to deliver ZFNs in seeds for targeted genome engineering.- Gene editing in channel catfish via double electroporation of zinc finger nucleases.- Delivery of mtZFNs into early mouse embryos.- Stereotaxic surgery and viral-delivery of zinc finger epigenetic editing tools in rodent brain.- In-vivo applications of cell-penetrating zinc finger transcription factors.- Manufacturing and delivering genome-editing proteins.
Riassunto
This detailed volume explores the notable progress in the field of zinc finger proteins (ZFP) study through widely used methods and protocols involving their biological functions and applications. Beginning with a section on the basic biology of ZFPs and design and applications of custom ZFPs, the book continues by covering methods for the evaluation and prevention of ZFN-mediated cytotoxicity as well as a collection of the representative methods of ZFN delivery. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Zinc Finger Proteins: Methods and Protocols aims to prompt both the understanding of ZFP biology and the development of next-generation ZFP therapeutics.
