PMID- 33612827
OWN - NLM
STAT- MEDLINE
DCOM- 20211027
LR  - 20231107
IS  - 1476-5462 (Electronic)
IS  - 0969-7128 (Print)
IS  - 0969-7128 (Linking)
VI  - 28
IP  - 7-8
DP  - 2021 Aug
TI  - Use of CRISPR/Cas9-mediated disruption of CNS cell type genes to profile 
      transduction of AAV by neonatal intracerebroventricular delivery in mice.
PG  - 456-468
LID - 10.1038/s41434-021-00223-3 [doi]
AB  - Adeno-associated virus (AAV) transduction efficiency and tropism are 
      conventionally determined by high expression of a fluorescent reporter gene. 
      Emerging data has suggested that such conventional methods may underestimate AAV 
      transduction for cells in which reporter expression from AAV vectors is 
      undetectable. To explore an alternative method that captures AAV transduction in 
      cells in which low expression of a cargo is sufficient for the intended activity, 
      we sought after CRISPR/Cas9-mediated gene disruption. In this study, we use AAV 
      to deliver CRISPR/guide RNA designed to abolish the genes NeuN, GFAP, or MOG 
      expressed specifically in neurons, astrocytes, or oligodendrocytes respectively 
      in the central nervous system (CNS) of mice. Abrogated expression of these 
      cell-type-specific genes can be measured biochemically in CNS subregions and 
      provides quantitative assessment of AAV transduction in these CNS cell types. By 
      using this method, we compared CNS transduction of AAV9, AAV-PHP.B, and 
      AAV-PHP.eB delivered via intracerebroventricular injection (ICV) in neonatal 
      mice. We found both AAV-PHP.B and AAV-PHP.eB resulted in marked disruption of the 
      NeuN gene by CRISPR/Cas9, significantly greater than AAV9 in several brain 
      regions and spinal cord. In contrast, only modest disruption of the GFAP gene and 
      the MOG gene was observed by all three AAV variants. Since the procedure of ICV 
      circumvents the blood-brain barrier, our data suggests that, independent of their 
      ability to cross the blood-brain barrier, AAV-PHP.B variants also exhibit 
      remarkably improved neuronal transduction in the CNS. We anticipate this approach 
      will facilitate profiling of AAV cellular tropism in murine CNS.
CI  - (c) 2021. The Author(s), under exclusive licence to Springer Nature Limited part of 
      Springer Nature.
FAU - Torregrosa, Tess
AU  - Torregrosa T
AUID- ORCID: 0000-0001-9870-0993
AD  - Biogen, Cambridge, MA, USA.
FAU - Lehman, Sydney
AU  - Lehman S
AD  - Biogen, Cambridge, MA, USA.
FAU - Hana, Sam
AU  - Hana S
AD  - Biogen, Cambridge, MA, USA.
FAU - Marsh, Galina
AU  - Marsh G
AD  - Biogen, Cambridge, MA, USA.
FAU - Xu, Shanqin
AU  - Xu S
AD  - Biogen, Cambridge, MA, USA.
FAU - Koszka, Kathryn
AU  - Koszka K
AD  - Biogen, Cambridge, MA, USA.
FAU - Mastrangelo, Nicole
AU  - Mastrangelo N
AD  - Biogen, Cambridge, MA, USA.
FAU - McCampbell, Alexander
AU  - McCampbell A
AD  - Biogen, Cambridge, MA, USA.
FAU - Henderson, Christopher E
AU  - Henderson CE
AD  - Biogen, Cambridge, MA, USA.
FAU - Lo, Shih-Ching
AU  - Lo SC
AUID- ORCID: 0000-0002-5187-0730
AD  - Biogen, Cambridge, MA, USA. joyce.lo@biogen.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210222
PL  - England
TA  - Gene Ther
JT  - Gene therapy
JID - 9421525
SB  - IM
MH  - Animals
MH  - CRISPR-Cas Systems
MH  - Central Nervous System
MH  - *Dependovirus/genetics
MH  - Gene Transfer Techniques
MH  - *Genetic Vectors/genetics
MH  - Mice
MH  - Neurons
MH  - Transduction, Genetic
PMC - PMC8376643
COIS- The authors declare that they have no conflict of interest.
EDAT- 2021/02/23 06:00
MHDA- 2021/10/28 06:00
PMCR- 2021/02/22
CRDT- 2021/02/22 05:50
PHST- 2020/07/03 00:00 [received]
PHST- 2021/01/15 00:00 [accepted]
PHST- 2020/12/01 00:00 [revised]
PHST- 2021/02/23 06:00 [pubmed]
PHST- 2021/10/28 06:00 [medline]
PHST- 2021/02/22 05:50 [entrez]
PHST- 2021/02/22 00:00 [pmc-release]
AID - 10.1038/s41434-021-00223-3 [pii]
AID - 223 [pii]
AID - 10.1038/s41434-021-00223-3 [doi]
PST - ppublish
SO  - Gene Ther. 2021 Aug;28(7-8):456-468. doi: 10.1038/s41434-021-00223-3. Epub 2021 
      Feb 22.