Efficient Mutagenesis by Cas9 Protein-Mediated Oligonucleotide Insertion and Large-Scale Assessment of Single-Guide RNAs

James A. Gagnon; Eivind Valen; Summer B. Thyme; Peng Huang; Laila Ahkmetova; Andrea Pauli; Tessa G. Montague; Steven Zimmerman; Constance Richter; Alexander F. Schier

doi:10.1371/journal.pone.0098186

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Peer-reviewed

Research Article

Efficient Mutagenesis by Cas9 Protein-Mediated Oligonucleotide Insertion and Large-Scale Assessment of Single-Guide RNAs

James A. Gagnon ,

* E-mail: james.gagnon@gmail.com (JAG); schier@fas.harvard.edu (AFS)

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Eivind Valen,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Summer B. Thyme,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Peng Huang,

Affiliations Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America, Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada
⨯
Laila Ahkmetova,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Andrea Pauli,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Tessa G. Montague,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Steven Zimmerman,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Constance Richter,

Affiliation Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America
⨯
Alexander F. Schier

* E-mail: james.gagnon@gmail.com (JAG); schier@fas.harvard.edu (AFS)

Affiliations Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, United States of America, Center for Brain Science, Harvard University, Cambridge, Massachusetts, United States of America, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts, United States of America, Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, United States of America
⨯

Efficient Mutagenesis by Cas9 Protein-Mediated Oligonucleotide Insertion and Large-Scale Assessment of Single-Guide RNAs

James A. Gagnon,
Eivind Valen,
Summer B. Thyme,
Peng Huang,
Laila Ahkmetova,
Andrea Pauli,
Tessa G. Montague,
Steven Zimmerman,
Constance Richter,
Alexander F. Schier

Published: May 29, 2014
https://doi.org/10.1371/journal.pone.0098186

Reader Comments (8)

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25 Nov 2014
18:04 GMT
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eomesA, oct4, mezzo oligo sequences? Posted by JamesGagnon on 25 Nov 2014 at 18:04 GMT
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25 Nov 2014
17:40 GMT
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optimize PCR for somatic insertion rates? Posted by JamesGagnon on 25 Nov 2014 at 17:40 GMT
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25 Nov 2014
17:37 GMT
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Orientation of the stop codon cassette oligo? Posted by JamesGagnon on 25 Nov 2014 at 17:37 GMT
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25 Nov 2014
17:36 GMT
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Why do some guide RNAs fail? Posted by JamesGagnon on 25 Nov 2014 at 17:36 GMT
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25 Nov 2014
16:35 GMT
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Buffer sequences in stop codon cassette? Posted by JamesGagnon on 25 Nov 2014 at 16:35 GMT
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25 Nov 2014
16:33 GMT
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Worry about SNPs? Posted by JamesGagnon on 25 Nov 2014 at 16:33 GMT
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25 Nov 2014
16:30 GMT
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Commercially Available Protein? Posted by JamesGagnon on 25 Nov 2014 at 16:30 GMT
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25 Nov 2014
16:25 GMT
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Oligo Insertion Rates? Posted by JamesGagnon on 25 Nov 2014 at 16:25 GMT

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Genomics
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Oligonucleotides
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