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High levels of AAV ...
High levels of AAV vector integration into CRISPR-induced DNA breaks
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- Hanlon, Killian S. (author)
- Harvard Med Sch, Dept Neurobiol, Boston, MA 02115 USA;Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA
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- Kleinstiver, Benjamin P. (author)
- Massachusetts Gen Hosp, Ctr Genom Med, Boston, MA 02114 USA;Massachusetts Gen Hosp, Dept Pathol, Boston, MA 02114 USA;Harvard Med Sch, Dept Pathol, Boston, MA 02115 USA
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- Garcia, Sara P. (author)
- Harvard Med Sch, Dept Pathol, Boston, MA 02115 USA;Massachusetts Gen Hosp, Mol Pathol Unit, Charlestown, MA 02129 USA;Massachusetts Gen Hosp, Ctr Canc Res & Ctr Computat & Integrat Biol, Charlestown, MA USA
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- Zaborowski, Mikolaj P. (author)
- Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA;Harvard Med Sch, Program Neurosci, Boston, MA 02115 USA;Poznan Univ Med Sci, Dept Gynecol Obstet & Gynecol Oncol, Div Gynecol Oncol, PL-60535 Poznan, Poland
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- Volak, Adrienn (author)
- Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA;Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
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- Spirig, Stefan E. (author)
- Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
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- Muller, Alissa (author)
- Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
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- Sousa, Alexander A. (author)
- Massachusetts Gen Hosp, Mol Pathol Unit, Charlestown, MA 02129 USA;Massachusetts Gen Hosp, Ctr Canc Res & Ctr Computat & Integrat Biol, Charlestown, MA USA
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- Tsai, Shengdar Q. (author)
- St Jude Childrens Res Hosp, Dept Hematol, 332 N Lauderdale St, Memphis, TN 38105 USA
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- Bengtsson, Niclas E. (author)
- Univ Washington, Dept Neurol, Seattle, WA 98195 USA
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- Lööv, Camilla, 1982- (author)
- Uppsala universitet,Geriatrik
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- Ingelsson, Martin (author)
- Uppsala universitet,Geriatrik
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- Chamberlain, Jeffrey S. (author)
- Univ Washington, Dept Neurol, Seattle, WA 98195 USA
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- Corey, David P. (author)
- Harvard Med Sch, Dept Neurobiol, Boston, MA 02115 USA
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- Aryee, Martin J. (author)
- Harvard Med Sch, Dept Pathol, Boston, MA 02115 USA;Massachusetts Gen Hosp, Mol Pathol Unit, Charlestown, MA 02129 USA;Massachusetts Gen Hosp, Ctr Canc Res & Ctr Computat & Integrat Biol, Charlestown, MA USA;Harvard TH Chan Sch Publ Hlth, Dept Biostat, Boston, MA USA
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- Joung, J. Keith (author)
- Harvard Med Sch, Dept Pathol, Boston, MA 02115 USA;Massachusetts Gen Hosp, Mol Pathol Unit, Charlestown, MA 02129 USA;Massachusetts Gen Hosp, Ctr Canc Res & Ctr Computat & Integrat Biol, Charlestown, MA USA
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- Breakefield, Xandra O. (author)
- Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA;Harvard Med Sch, Program Neurosci, Boston, MA 02115 USA
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- Maguire, Casey A. (author)
- Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA;Harvard Med Sch, Program Neurosci, Boston, MA 02115 USA
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- György, Bence (author)
- Harvard Med Sch, Dept Neurobiol, Boston, MA 02115 USA;Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA;Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
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(creator_code:org_t)
- 2019-09-30
- 2019
- English.
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In: Nature Communications. - : NATURE PUBLISHING GROUP. - 2041-1723. ; 10
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Abstract
Subject headings
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- Adeno-associated virus (AAV) vectors have shown promising results in preclinical models, but the genomic consequences of transduction with AAV vectors encoding CRISPR-Cas nucleases is still being examined. In this study, we observe high levels of AAV integration (up to 47%) into Cas9-induced double-strand breaks (DSBs) in therapeutically relevant genes in cultured murine neurons, mouse brain, muscle and cochlea. Genome-wide AAV mapping in mouse brain shows no overall increase of AAV integration except at the CRISPR/Cas9 target site. To allow detailed characterization of integration events we engineer a miniature AAV encoding a 465 bp lambda bacteriophage DNA (AAV-lambda 465), enabling sequencing of the entire integrated vector genome. The integration profile of AAV-465 lambda in cultured cells display both full-length and fragmented AAV genomes at Cas9 on-target sites. Our data indicate that AAV integration should be recognized as a common outcome for applications that utilize AAV for genome editing.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Hanlon, Killian ...
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Kleinstiver, Ben ...
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Garcia, Sara P.
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Zaborowski, Miko ...
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Volak, Adrienn
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Spirig, Stefan E ...
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show more...
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Muller, Alissa
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Sousa, Alexander ...
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Tsai, Shengdar Q ...
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Bengtsson, Nicla ...
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Lööv, Camilla, 1 ...
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Ingelsson, Marti ...
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Chamberlain, Jef ...
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Corey, David P.
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Aryee, Martin J.
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Joung, J. Keith
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Breakefield, Xan ...
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Maguire, Casey A ...
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György, Bence
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- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Medical Biotechn ...
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and Medical Biotechn ...
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Nature Communica ...
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Uppsala University