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High levels of AAV vector integration into CRISPR-induced DNA breaks

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
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
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
Volak, Adrienn (author)
Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA;Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
Spirig, Stefan E. (author)
Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
Muller, Alissa (author)
Inst Mol & Clin Ophthalmol Basel, CH-4031 Basel, Switzerland
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
Tsai, Shengdar Q. (author)
St Jude Childrens Res Hosp, Dept Hematol, 332 N Lauderdale St, Memphis, TN 38105 USA
Bengtsson, Niclas E. (author)
Univ Washington, Dept Neurol, Seattle, WA 98195 USA
Lööv, Camilla, 1982- (author)
Uppsala universitet,Geriatrik
Ingelsson, Martin (author)
Uppsala universitet,Geriatrik
Chamberlain, Jeffrey S. (author)
Univ Washington, Dept Neurol, Seattle, WA 98195 USA
Corey, David P. (author)
Harvard Med Sch, Dept Neurobiol, Boston, MA 02115 USA
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
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
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
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
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.
In: Nature Communications. - : NATURE PUBLISHING GROUP. - 2041-1723. ; 10
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  • Journal article (peer-reviewed)
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)

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