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Dynamics of CRISPR-Cas9 genome interrogation in living cells

Genome editing with a Cas9 scalpelThe Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living c... Full description

Journal Title: Science November 13, 2015, Vol.350(6262), pp.823-826
Main Author: Knight, Spencer
Other Authors: Xie, Liangqi , Deng, Wulan , Guglielmi, Benjamin , Witkowsky, Lea , Bosanac, Lana , Zhang, Elisa , El Beheiry, Mohamed , Masson, Jean-Baptiste , Dahan, Maxime , Liu, Zhe , Doudna, Jennifer , Tjian, Robert
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 0036-8075 ; DOI: 10.1126/science.aac6572
Link: http://search.proquest.com/docview/1825565985/?pq-origsite=primo
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title: Dynamics of CRISPR-Cas9 genome interrogation in living cells
format: Article
creator:
  • Knight, Spencer
  • Xie, Liangqi
  • Deng, Wulan
  • Guglielmi, Benjamin
  • Witkowsky, Lea
  • Bosanac, Lana
  • Zhang, Elisa
  • El Beheiry, Mohamed
  • Masson, Jean-Baptiste
  • Dahan, Maxime
  • Liu, Zhe
  • Doudna, Jennifer
  • Tjian, Robert
subjects:
  • Genomes
  • Cells (Biology)
  • Bacteria
  • Ribonucleic Acids
  • Searching
  • Diffusion
  • Editing
  • Binding
  • Miscellaneous Sciences (So)
  • (An)
ispartof: Science, November 13, 2015, Vol.350(6262), pp.823-826
description: Genome editing with a Cas9 scalpelThe Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living cells. Cas9 searches the genome for its target sites using rapid threedimensional diffusion. It spends very little time binding to off-target sites, which explains the high accuracy of the CRISPRCas9 editing machine.Science, this issue p. 823 The RNA-guided CRISPR-associated protein Cas9 is used for genome editing, transcriptional modulation, and live-cell imaging. Cas9-guide RNA complexes recognize and cleave double-stranded DNA sequences on the basis of 20-nucleotide RNA-DNA complementarity, but the mechanism of target searching in mammalian cells is unknown. Here, we use single-particle tracking to visualize diffusion and chromatin binding of Cas9 in living cells. We...
language: eng
source:
identifier: ISSN: 0036-8075 ; DOI: 10.1126/science.aac6572
fulltext: no_fulltext
issn:
  • 00368075
  • 0036-8075
url: Link


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titleDynamics of CRISPR-Cas9 genome interrogation in living cells
creatorKnight, Spencer ; Xie, Liangqi ; Deng, Wulan ; Guglielmi, Benjamin ; Witkowsky, Lea ; Bosanac, Lana ; Zhang, Elisa ; El Beheiry, Mohamed ; Masson, Jean-Baptiste ; Dahan, Maxime ; Liu, Zhe ; Doudna, Jennifer ; Tjian, Robert
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ispartofScience, November 13, 2015, Vol.350(6262), pp.823-826
identifierISSN: 0036-8075 ; DOI: 10.1126/science.aac6572
subjectGenomes ; Cells (Biology) ; Bacteria ; Ribonucleic Acids ; Searching ; Diffusion ; Editing ; Binding ; Miscellaneous Sciences (So) ; (An)
descriptionGenome editing with a Cas9 scalpelThe Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living cells. Cas9 searches the genome for its target sites using rapid threedimensional diffusion. It spends very little time binding to off-target sites, which explains the high accuracy of the CRISPRCas9 editing machine.Science, this issue p. 823 The RNA-guided CRISPR-associated protein Cas9 is used for genome editing, transcriptional modulation, and live-cell imaging. Cas9-guide RNA complexes recognize and cleave double-stranded DNA sequences on the basis of 20-nucleotide RNA-DNA complementarity, but the mechanism of target searching in mammalian cells is unknown. Here, we use single-particle tracking to visualize diffusion and chromatin binding of Cas9 in living cells. We...
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titleDynamics of CRISPR-Cas9 genome interrogation in living cells
descriptionGenome editing with a Cas9 scalpelThe Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living cells. Cas9 searches the genome for its target sites using rapid threedimensional diffusion. It spends very little time binding to off-target sites, which explains the high accuracy of the CRISPRCas9 editing machine.Science, this issue p. 823 The RNA-guided CRISPR-associated protein Cas9 is used for genome editing, transcriptional modulation, and live-cell imaging. Cas9-guide RNA complexes recognize and cleave double-stranded DNA sequences on the basis of 20-nucleotide RNA-DNA complementarity, but the mechanism of target searching in mammalian cells is unknown. Here, we use single-particle tracking to visualize diffusion and chromatin binding of Cas9 in living cells. We...
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abstractGenome editing with a Cas9 scalpelThe Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living cells. Cas9 searches the genome for its target sites using rapid threedimensional diffusion. It spends very little time binding to off-target sites, which explains the high accuracy of the CRISPRCas9 editing machine.Science, this issue p. 823 The RNA-guided CRISPR-associated protein Cas9 is used for genome editing, transcriptional modulation, and live-cell imaging. Cas9-guide RNA complexes recognize and cleave double-stranded DNA sequences on the basis of 20-nucleotide RNA-DNA complementarity, but the mechanism of target searching in mammalian cells is unknown. Here, we use single-particle tracking to visualize diffusion and chromatin binding of Cas9 in living cells. We...
doi10.1126/science.aac6572
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date2015-11-13