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High levels of AAV vector integration into CRISPR-induced DNA breaks
Harvard Med Sch, Dept Neurobiol, Boston, MA 02115 USA;Massachusetts Gen Hosp, Dept Neurol, Mol Neurogenet Unit, Charlestown, MA 02129 USA.
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.
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.
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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2019 (engelsk)Inngår i: Nature Communications, ISSN 2041-1723, E-ISSN 2041-1723, Vol. 10, artikkel-id 4439Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

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.

sted, utgiver, år, opplag, sider
NATURE PUBLISHING GROUP , 2019. Vol. 10, artikkel-id 4439
HSV kategori
Identifikatorer
URN: urn:nbn:se:uu:diva-395832DOI: 10.1038/s41467-019-12449-2ISI: 000488235000009PubMedID: 31570731OAI: oai:DiVA.org:uu-395832DiVA, id: diva2:1365615
Tilgjengelig fra: 2019-10-25 Laget: 2019-10-25 Sist oppdatert: 2019-10-25bibliografisk kontrollert

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