Modern Prometheus

Filename: modern-prometheus.pdf
ISBN: 9781316780985
Release Date: 2016-09-30
Number of pages:
Author: James Kozubek
Publisher: Cambridge University Press

Download and read online Modern Prometheus in PDF and EPUB Would you change your genes if you could? As we confront the 'industrial revolution of the genome', the recent discoveries of Crispr-Cas9 technologies are offering, for the first time, cheap and effective methods for editing the human genome. This opens up startling new opportunities as well as significant ethical uncertainty. Tracing events across a fifty-year period, from the first gene splicing techniques to the present day, this is the story of gene editing - the science, the impact and the potential. Kozubek weaves together the fascinating stories of many of the scientists involved in the development of gene editing technology. Along the way, he demystifies how the technology really works and provides vivid and thought-provoking reflections on the continuing ethical debate. Ultimately, Kozubek places the debate in its historical and scientific context to consider both what drives scientific discovery and the implications of the 'commodification' of life.


Crispr Cas A Laboratory Manual

Filename: crispr-cas-a-laboratory-manual.pdf
ISBN: 1621821315
Release Date: 2016-03-31
Number of pages: 182
Author: Jennifer Doudna
Publisher:

Download and read online Crispr Cas A Laboratory Manual in PDF and EPUB The development of CRISPR-Cas technology is revolutionizing biology. Based on machinery bacteria use to target foreign DNA, these powerful techniques allow investigators to edit DNA sequences and modulate gene expression more rapidly and accurately than ever before. Featuring contributions from leading figures in the CRISPR-Cas field, this laboratory manual presents a state-of-the-art guide to the technology. It includes step-by-step protocols for applying CRISPR-Cas-based techniques in various systems, including yeast, zebrafish, Drosophila, mice, and cultured cells (e.g., human pluripotent stem cells). The contributors cover web-based tools and approaches for designing guide RNAs that precisely target genes of interest, methods for preparing and delivering CRISPR-Cas reagents into cells, and ways to screen for cells that harbor the desired genetic changes. Strategies for optimizing CRISPR-Cas in each system-especially for minimizing off-target effects-are also provided. Authors also describe other applications of the CRISPR-Cas system, including its use for regulating genome activation and repression, and discuss the development of next-generation CRISPR-Cas tools. The book is thus an essential laboratory resource for all cell, molecular, and developmental biologists, as well as biochemists, geneticists, and all who seek to expand their biotechnology toolkits.


CRISPR

Filename: crispr.pdf
ISBN: 1493954288
Release Date: 2016-10-09
Number of pages: 366
Author: Magnus Lundgren
Publisher: Humana Press

Download and read online CRISPR in PDF and EPUB This volume presents a list of cutting-edge protocols for the study of CRISPR-Cas defense systems and their applications at the genomic, genetic, biochemical and structural levels. CRISPR: Methods and Protocols guides readers through techniques that have been developed specifically for the analysis of CRISPR-Cas and techniques adapted from standard protocols of DNA, RNA and protein biology. Written in the highly successful Methods in Molecular Biology series format, 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 cutting-edge, CRISPR: Methods and Protocols provides a broad list of tools and techniques to study the interdisciplinary aspects of the prokaryotic CRISPR-Cas defense systems.


The Use of CRISPR cas9 ZFNs TALENs in Generating Site Specific Genome Alterations

Filename: the-use-of-crispr-cas9-zfns-talens-in-generating-site-specific-genome-alterations.pdf
ISBN: 9780128013342
Release Date: 2014-11-04
Number of pages: 570
Author:
Publisher: Academic Press

Download and read online The Use of CRISPR cas9 ZFNs TALENs in Generating Site Specific Genome Alterations in PDF and EPUB This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers recent research and methods development for changing the DNA sequence within the genomes of cells and organisms. Focusing on enzymes that generate double-strand breaks in DNA, the chapters describe use of molecular tools to introduce or delete genetic information at specific sites in the genomes of animal, plant and bacterial cells. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in biomineralization science Contains sections on such topics as genome editing, genome engineering, CRISPR, Cas9, TALEN and zinc finger nuclease


CRISPR Cas Systems

Filename: crispr-cas-systems.pdf
ISBN: 9783642346576
Release Date: 2012-12-13
Number of pages: 302
Author: Rodolphe Barrangou
Publisher: Springer Science & Business Media

Download and read online CRISPR Cas Systems in PDF and EPUB CRISPR/Cas is a recently described defense system that protects bacteria and archaea against invasion by mobile genetic elements such as viruses and plasmids. A wide spectrum of distinct CRISPR/Cas systems has been identified in at least half of the available prokaryotic genomes. On-going structural and functional analyses have resulted in a far greater insight into the functions and possible applications of these systems, although many secrets remain to be discovered. In this book, experts summarize the state of the art in this exciting field.



The Use of CRISPR cas9 ZFNs TALENs in Generating Site Specific Genome Alterations

Filename: the-use-of-crispr-cas9-zfns-talens-in-generating-site-specific-genome-alterations.pdf
ISBN: 9780128013342
Release Date: 2014-11-04
Number of pages: 570
Author:
Publisher: Academic Press

Download and read online The Use of CRISPR cas9 ZFNs TALENs in Generating Site Specific Genome Alterations in PDF and EPUB This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers recent research and methods development for changing the DNA sequence within the genomes of cells and organisms. Focusing on enzymes that generate double-strand breaks in DNA, the chapters describe use of molecular tools to introduce or delete genetic information at specific sites in the genomes of animal, plant and bacterial cells. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers research methods in biomineralization science Contains sections on such topics as genome editing, genome engineering, CRISPR, Cas9, TALEN and zinc finger nuclease


A Crack in Creation

Filename: a-crack-in-creation.pdf
ISBN: 9780544716964
Release Date: 2017-06-13
Number of pages: 320
Author: Jennifer A. Doudna
Publisher: Houghton Mifflin Harcourt

Download and read online A Crack in Creation in PDF and EPUB A trailblazing biologist grapples with her role in the biggest scientific discovery of our era: a cheap, easy way of rewriting genetic code, with nearly limitless promise and peril. Not since the atomic bomb has a technology so alarmed its inventors that they warned the world about its use. Not, that is, until the spring of 2015, when biologist Jennifer Doudna called for a worldwide moratorium on the use of the new gene-editing tool CRISPR—a revolutionary new technology that she helped create—to make heritable changes in human embryos. The cheapest, simplest, most effective way of manipulating DNA ever known, CRISPR may well give us the cure to HIV, genetic diseases, and some cancers, and will help address the world’s hunger crisis. Yet even the tiniest changes to DNA could have myriad, unforeseeable consequences—to say nothing of the ethical and societal repercussions of intentionally mutating embryos to create “better” humans. Writing with fellow researcher Samuel Sternberg, Doudna shares the thrilling story of her discovery, and passionately argues that enormous responsibility comes with the ability to rewrite the code of life. With CRISPR, she shows, we have effectively taken control of evolution. What will we do with this unfathomable power?


CRISPR Cas9 mediated Protein Tagging in Human Cells for RESOLFT Nanoscopy and the Analysis of Mitochondrial Prohibitins

Filename: crispr-cas9-mediated-protein-tagging-in-human-cells-for-resolft-nanoscopy-and-the-analysis-of-mitochondrial-prohibitins.pdf
ISBN: OCLC:965869580
Release Date: 2016
Number of pages:
Author: Michael Ratz
Publisher:

Download and read online CRISPR Cas9 mediated Protein Tagging in Human Cells for RESOLFT Nanoscopy and the Analysis of Mitochondrial Prohibitins in PDF and EPUB Ectopic overexpression of fluorescent fusion proteins for live cell imaging studies often leads to a multitude of artefacts, but protein expression at endogenous levels in mammalian cells was difficult to achieve so far. To avoid common problems associated with overexpression, this study used the CRISPR-Cas9 genome engineering system for site-specific endogenous protein tagging in human cells. First, a general workflow for genome editing was established and then applied to generate heterozygous and homozygous human knock-in cells that express a fluorescent fusion from a genomic locus. Three...


Development of Whole genome CRISPR Cas9 Functional Screens for the Discovery of the TH 302 Sensitivity Genes

Filename: development-of-whole-genome-crispr-cas9-functional-screens-for-the-discovery-of-the-th-302-sensitivity-genes.pdf
ISBN: OCLC:962372563
Release Date: 2016
Number of pages: 350
Author: Indumati Sharma
Publisher:

Download and read online Development of Whole genome CRISPR Cas9 Functional Screens for the Discovery of the TH 302 Sensitivity Genes in PDF and EPUB Hypoxia, a central pathophysiological feature of many tumours, has long been considered as a potential target for solid tumour-selective therapy. In addition to solid tumours, recent evidence indicates the role of hypoxic bone marrow in advancing leukaemia and thus suggesting that hypoxic bone marrow niche may also be considered for targeted therapies in leukaemic patients. Hypoxia-activated prodrugs (HAPs) contain a bioreductive cytotoxic moiety which is selectively activated in hypoxic tumour cells. The clinical development of the HAP TH-302 has been challenging. This thesis aimed to develop the methodologies for harnessing the recent, RNA guided CRISPR (clustered regularly interspaced short palindromic repeats)-associated Cas9 nuclease (CRISPR-Cas9) for performing high-throughput genome-scale knockout (GeCKO) to identify the genetic sensitivity determinants of TH-302. Production of high titer lentiviruses (LV) using sucrose gradient centrifugation improved the transduction efficiency of Cas9 expressing pseudohaploid chronic myeloid leukaemia cell line KBM7 (KBM7-Cas9). Using high titer LV, a GeCKO library targeting 19,050 genes with unique 122,411 guide RNA sequences was produced at an MOI of ~0.25 in KBM7-Cas9 cells. The ensuing KBM7-Cas9-GeCKO library was screened with TH-302 and its active metabolite Br-IPM, to identify genetic sensitivity determinants. Drug screening methodology was optimised where low concentrations yet repeated cycles of drug exposure prevented the surviving fraction from undergoing tremendous genetic bottleneck, thus retaining a significant proportion of the library with adequate representation. To determine the functioning of the current bioinformatics pipeline, MAGeCK -VISPR analysis of the IIlumina HiSeq data from previously performed pilot-study with 6-thioguanine (6-TG) in a small KBM7-Cas9-GeCKO library, was carried out. The high-ranking hits identified several genes with known roles in 6-thioguanine sensitivity (NUDT5, MLH1, MSH6, PMS2, HPRT1). The end-to-end pipeline of CRISPR-Cas9 functional screens at ACSRC was established to answer the important long term objective of the project, i.e., to identify the determinants of sensitivity to TH-302.



A CRISPR Cas9 Approach for in Vivo Tissue specific Genetic Disruptions of Cell Cycle Regulators in Zebrafish Danio Rerio Eyes

Filename: a-crispr-cas9-approach-for-in-vivo-tissue-specific-genetic-disruptions-of-cell-cycle-regulators-in-zebrafish-danio-rerio-eyes.pdf
ISBN: OCLC:953209160
Release Date: 2016
Number of pages: 59
Author: Bailey L. Simon
Publisher:

Download and read online A CRISPR Cas9 Approach for in Vivo Tissue specific Genetic Disruptions of Cell Cycle Regulators in Zebrafish Danio Rerio Eyes in PDF and EPUB



Genome Editing with the CRISPR Cas9 System

Filename: genome-editing-with-the-crispr-cas9-system.pdf
ISBN: 3659851701
Release Date: 2016-03-12
Number of pages: 64
Author: Paris Roidos
Publisher:

Download and read online Genome Editing with the CRISPR Cas9 System in PDF and EPUB


Targeted Genome Editing Using Site Specific Nucleases

Filename: targeted-genome-editing-using-site-specific-nucleases.pdf
ISBN: 9784431552277
Release Date: 2015-01-05
Number of pages: 205
Author: Takashi Yamamoto
Publisher: Springer

Download and read online Targeted Genome Editing Using Site Specific Nucleases in PDF and EPUB This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.