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3549250 Doudna, Jennifer; Mali, Prashant:
CRISPR-Cas: A Laboratory Manual
  Preis:   € 98,00

Einband: Psperback
Auflage: 1.Auflage
Verlag: Cold Spring Harbor Laboratory Press New York
Erscheinungsdatum: 03/2016
Seiten: 450 pages (approx.)
Abbildungen: illustrations (4C & B&W), index

ISBN-10: 1-62182-131-5   
ISBN-13: 9781621821311


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Beschreibung
CRISPR-Cas-based techniques are revolutionizing the way geneticists and molecular biologists modify DNA sequences and modulate gene expression in cells and organisms. This laboratory manual presents step-by-step protocols for applying this cutting-edge technology to any system of interest. Contributors describe approaches for designing guide RNAs that precisely target the gene of interest, delivering CRISPR-Cas reagents into cells (e.g., using AAV vectors), and selecting cells that harbor the desired changes. Strategies for scaling and multiplexing these procedures are also described, as are methods for optimizing CRISPR-Cas systems in common species and cell lines, from yeast to human.
Inhalt
Contents

(Preliminary)
1 Overview of CRISPR-Cas9 Biology
Hannah K Ratner, Timothy R Sampson, and David S Weiss
(Emory University School of Medicine and Emory Vaccine Center)
2 Prediction and Validation of Native and Engineered Cas9 Guide Sequences
Alexandra E. Briner, Emily D. Henriksen, and Rodolphe Barrangou
(North Carolina State University)
3 Characterization of Cas9-gRNA Orthologs
Jonathan L. Braff, Stephanie J. Yaung, Kevin M. Esvelt, and George M. Church
(Harvard Medical School)
4 Detection of Single Nucleotide Substitutions Induced by Genome Editing Using Digital PCR
Yuichiro Miyaoka, Amanda H. Chan, and Bruce R. Conklin
(Gladstone Institute, University of California San Francisco)
5 Genome Editing in Human Pluripotent Stem Cells
Cory Smith, Zhaohui Ye, and Linzhao Cheng
(Johns Hopkins University School of Medicine)
6 CRISPR-Cas9 Genome Engineering of Saccharomyces cerevisiae Cells
Owen W. Ryan, Snigdha Poddar, and Jamie H. D. Cate
(University of California Berkeley)
7 CRISPR Technology for Genome Activation and Repression in Mammalian Cells
Dan Du and Lei S. Qi
(Stanford University)
8 Cas9-mediated Genome Engineering in Drosophila melanogaster
Benjamin Housden, Yanhui Hu, and Norbert Perrimon
(Harvard Medical School)
9 Optimized CRISPR-Cas9 System for Genome Editing in Zebrafish
Charles E Vejnar, Miguel Moreno-Mateos, Daniel Cifuentes, Ariel A Bazzini, and Antonio J.Giraldez
(Yale University School of Medicine)
10 AAV-mediated delivery of CRISPR-Cas Systems for Genome Engineering in Mammalian Cells
Thomas Gaj and David V. Schaffer
(University of California Berkeley)
11 Large-scale Single-guide RNA Library Construction and Use for Genetic Screens
Tim Wang (Massachusetts Institute of Technology), Eric S. Lander (Broad Institute of MIT and Harvard Medical School), and David M. Sabatini (Massachusetts Institute of Technology)
12 The Generation of Genetically Modified Mice Using the CRISPR-Cas9 Genome Editing System
Adam Williams (University of Connecticut Health Center), Jorge Henao-Mejia (University of Pennsylvania Perelman School of Medicine), and Richard A. Flavell (HHMI/Yale University School of Medicine)
Index

Zielgruppe/Leser
Developmental Biology; Gene Therapy; Cell Biology; Genomics; Molecular Biology; Mouse Biology; Genetics; Biochemistry; Laboratory Techniques
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