PMID- 30863790
OWN - NLM
STAT- MEDLINE
DCOM- 20190418
LR  - 20240412
IS  - 2373-2822 (Electronic)
IS  - 2373-2822 (Linking)
VI  - 6
IP  - 1
DP  - 2019 Jan-Feb
TI  - A Neuron-Optimized CRISPR/dCas9 Activation System for Robust and Specific Gene 
      Regulation.
LID - 10.1523/ENEURO.0495-18.2019 [doi]
LID - ENEURO.0495-18.2019
AB  - CRISPR-based technology has provided new avenues to interrogate gene function, 
      but difficulties in transgene expression in post-mitotic neurons has delayed 
      incorporation of these tools in the central nervous system (CNS). Here, we 
      demonstrate a highly efficient, neuron-optimized dual lentiviral CRISPR-based 
      transcriptional activation (CRISPRa) system capable of robust, modular, and 
      tunable gene induction and multiplexed gene regulation across several primary 
      rodent neuron culture systems. CRISPRa targeting unique promoters in the complex 
      multi-transcript gene brain-derived neurotrophic factor (Bdnf) revealed both 
      transcript- and genome-level selectivity of this approach, in addition to 
      highlighting downstream transcriptional and physiological consequences of Bdnf 
      regulation. Finally, we illustrate that CRISPRa is highly efficient in vivo, 
      resulting in increased protein levels of a target gene in diverse brain 
      structures. Taken together, these results demonstrate that CRISPRa is an 
      efficient and selective method to study gene expression programs in brain health 
      and disease.
FAU - Savell, Katherine E
AU  - Savell KE
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Bach, Svitlana V
AU  - Bach SV
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Zipperly, Morgan E
AU  - Zipperly ME
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Revanna, Jasmin S
AU  - Revanna JS
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Goska, Nicholas A
AU  - Goska NA
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Tuscher, Jennifer J
AU  - Tuscher JJ
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Duke, Corey G
AU  - Duke CG
AUID- ORCID: 0000-0003-1102-9549
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Sultan, Faraz A
AU  - Sultan FA
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Burke, Julia N
AU  - Burke JN
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Williams, Derek
AU  - Williams D
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
FAU - Ianov, Lara
AU  - Ianov L
AD  - Civitan International Research Center, University of Alabama at Birmingham, 
      Birmingham, AL, 35294.
FAU - Day, Jeremy J
AU  - Day JJ
AUID- ORCID: 0000-0002-7361-3399
AD  - Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of 
      Alabama at Birmingham, Birmingham, AL, 35294.
AD  - Civitan International Research Center, University of Alabama at Birmingham, 
      Birmingham, AL, 35294.
LA  - eng
GR  - DP1 DA039650/DA/NIDA NIH HHS/United States
GR  - F32 DA041778/DA/NIDA NIH HHS/United States
GR  - R00 DA034681/DA/NIDA NIH HHS/United States
GR  - F32 MH112304/MH/NIMH NIH HHS/United States
GR  - F31 DA042514/DA/NIDA NIH HHS/United States
GR  - R01 MH114990/MH/NIMH NIH HHS/United States
GR  - T32 GM008361/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20190307
PL  - United States
TA  - eNeuro
JT  - eNeuro
JID - 101647362
RN  - 0 (Bdnf protein, rat)
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Cell Adhesion Molecules, Neuronal)
RN  - 0 (Extracellular Matrix Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Reelin Protein)
RN  - EC 3.4.21.- (Serine Endopeptidases)
SB  - IM
MH  - Animals
MH  - Brain/cytology/metabolism
MH  - Brain-Derived Neurotrophic Factor/genetics/metabolism
MH  - *CRISPR-Cas Systems
MH  - Cell Adhesion Molecules, Neuronal/metabolism
MH  - Cell Line, Tumor
MH  - Extracellular Matrix Proteins/metabolism
MH  - *Gene Expression Regulation
MH  - *Genetic Techniques
MH  - Male
MH  - Nerve Tissue Proteins/metabolism
MH  - Neurons/cytology/*metabolism
MH  - Primary Cell Culture
MH  - Random Allocation
MH  - Rats, Sprague-Dawley
MH  - Reelin Protein
MH  - Serine Endopeptidases/metabolism
MH  - Transcription, Genetic
MH  - Transcriptome
PMC - PMC6412672
OTO - NOTNLM
OT  - Bdnf
OT  - CRISPR
OT  - epigenetics
OT  - gene expression
OT  - transcription
EDAT- 2019/03/14 06:00
MHDA- 2019/04/19 06:00
PMCR- 2019/03/07
CRDT- 2019/03/14 06:00
PHST- 2018/12/18 00:00 [received]
PHST- 2019/01/21 00:00 [revised]
PHST- 2019/01/27 00:00 [accepted]
PHST- 2019/03/14 06:00 [entrez]
PHST- 2019/03/14 06:00 [pubmed]
PHST- 2019/04/19 06:00 [medline]
PHST- 2019/03/07 00:00 [pmc-release]
AID - eN-MNT-0495-18 [pii]
AID - 10.1523/ENEURO.0495-18.2019 [doi]
PST - epublish
SO  - eNeuro. 2019 Mar 7;6(1):ENEURO.0495-18.2019. doi: 10.1523/ENEURO.0495-18.2019. 
      eCollection 2019 Jan-Feb.