PMID- 27587996
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20160902
LR  - 20200929
IS  - 1662-5099 (Print)
IS  - 1662-5099 (Electronic)
IS  - 1662-5099 (Linking)
VI  - 9
DP  - 2016
TI  - The Development of a Viral Mediated CRISPR/Cas9 System with Doxycycline Dependent 
      gRNA Expression for Inducible In vitro and In vivo Genome Editing.
PG  - 70
LID - 10.3389/fnmol.2016.00070 [doi]
LID - 70
AB  - The RNA-guided Cas9 nuclease, from the type II prokaryotic Clustered Regularly 
      Interspersed Short Palindromic Repeats (CRISPR) adaptive immune system, has been 
      adapted and utilized by scientists to edit the genomes of eukaryotic cells. Here, 
      we report the development of a viral mediated CRISPR/Cas9 system that can be 
      rendered inducible utilizing doxycycline (Dox) and can be delivered to cells in 
      vitro and in vivo utilizing adeno-associated virus (AAV). Specifically, we 
      developed an inducible gRNA (gRNAi) AAV vector that is designed to express the 
      gRNA from a H1/TO promoter. This AAV vector is also designed to express the Tet 
      repressor (TetR) to regulate the expression of the gRNAi in a Dox dependent 
      manner. We show that H1/TO promoters of varying length and a U6/TO promoter can 
      edit DNA with similar efficiency in vitro, in a Dox dependent manner. We also 
      demonstrate that our inducible gRNAi vector can be used to edit the genomes of 
      neurons in vivo within the mouse brain in a Dox dependent manner. Genome editing 
      can be induced in vivo with this system by supplying animals Dox containing food 
      for as little as 1 day. This system might be cross compatible with many existing 
      S. pyogenes Cas9 systems (i.e., Cas9 mouse, CRISPRi, etc.), and therefore it 
      likely can be used to render these systems inducible as well.
FAU - de Solis, Christopher A
AU  - de Solis CA
AD  - School of Behavioral and Brain Sciences, Department of Molecular and Cell 
      Biology, University of Texas at Dallas Richardson, TX, USA.
FAU - Ho, Anthony
AU  - Ho A
AD  - School of Behavioral and Brain Sciences, Department of Molecular and Cell 
      Biology, University of Texas at Dallas Richardson, TX, USA.
FAU - Holehonnur, Roopashri
AU  - Holehonnur R
AD  - School of Behavioral and Brain Sciences, Department of Molecular and Cell 
      Biology, University of Texas at Dallas Richardson, TX, USA.
FAU - Ploski, Jonathan E
AU  - Ploski JE
AD  - School of Behavioral and Brain Sciences, Department of Molecular and Cell 
      Biology, University of Texas at Dallas Richardson, TX, USA.
LA  - eng
GR  - R21 MH109945/MH/NIMH NIH HHS/United States
PT  - Journal Article
DEP - 20160818
PL  - Switzerland
TA  - Front Mol Neurosci
JT  - Frontiers in molecular neuroscience
JID - 101477914
PMC - PMC4988984
OTO - NOTNLM
OT  - AAV vectors
OT  - CRISPR/Cas9
OT  - TET2
OT  - TRE3G
OT  - amygdala
OT  - doxycycline
OT  - genome editing
OT  - inducible
EDAT- 2016/09/03 06:00
MHDA- 2016/09/03 06:01
PMCR- 2016/01/01
CRDT- 2016/09/03 06:00
PHST- 2016/06/16 00:00 [received]
PHST- 2016/07/29 00:00 [accepted]
PHST- 2016/09/03 06:00 [entrez]
PHST- 2016/09/03 06:00 [pubmed]
PHST- 2016/09/03 06:01 [medline]
PHST- 2016/01/01 00:00 [pmc-release]
AID - 10.3389/fnmol.2016.00070 [doi]
PST - epublish
SO  - Front Mol Neurosci. 2016 Aug 18;9:70. doi: 10.3389/fnmol.2016.00070. eCollection 
      2016.