PMID- 23531602
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20130328
LR  - 20220809
IS  - 2162-2531 (Print)
IS  - 2162-2531 (Electronic)
IS  - 2162-2531 (Linking)
VI  - 2
IP  - 3
DP  - 2013 Mar 26
TI  - Voltage Preconditioning Allows Modulated Gene Expression in Neurons Using 
      PEI-complexed siRNA.
PG  - e82
LID - 10.1038/mtna.2013.10 [doi]
AB  - We present here a high efficiency, high viability siRNA-delivery method using a 
      voltage-controlled chemical transfection strategy to achieve modulated delivery 
      of polyethylenimine (PEI) complexed with siRNA in an in vitro culture of neuro2A 
      cells and neurons. Low voltage pulses were applied to adherent cells before the 
      administration of PEI-siRNA complexes. Live assays of neuro2a cells transfected 
      with fluorescently tagged siRNA showed an increase in transfection efficiency 
      from 62 +/- 14% to 98 +/- 3.8% (after -1 V). In primary hippocampal neurons, 
      transfection efficiencies were increased from 30 +/- 18% to 76 +/- 18% (after -1 V). 
      Negligible or low-level transfection was observed after preconditioning at higher 
      voltages, suggesting an inverse relationship with applied voltage. Experiments 
      with propidium iodide ruled out the role of electroporation in the transfection 
      of siRNAs suggesting an alternate electro-endocytotic mechanism. In addition, 
      image analysis of preconditioned and transfected cells demonstrates siRNA uptake 
      and loading that is tuned to preconditioning voltage levels. There is 
      approximately a fourfold increase in siRNA loading after preconditioning at -1 V 
      compared with the same at +/-2-3 V. Modulated gene expression is demonstrated in a 
      functional knockdown of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in 
      neuro2A cells using siRNA. Cell density and dendritic morphological changes are 
      also demonstrated in modulated knockdown of brain derived neurotrophic factor 
      (BDNF) in primary hippocampal neurons. The method reported here has potential 
      applications in the development of high-throughput screening systems for large 
      libraries of siRNA molecules involving difficult-to-transfect cells like 
      neurons.Molecular Therapy-Nucleic Acids (2013) 2, e82; doi:10.1038/mtna.2013.10; 
      published online 26 March 2013.
FAU - Sridharan, Arati
AU  - Sridharan A
FAU - Patel, Chetan
AU  - Patel C
FAU - Muthuswamy, Jit
AU  - Muthuswamy J
LA  - eng
GR  - F32 NS073422/NS/NINDS NIH HHS/United States
PT  - Journal Article
DEP - 20130326
PL  - United States
TA  - Mol Ther Nucleic Acids
JT  - Molecular therapy. Nucleic acids
JID - 101581621
PMC - PMC3615821
EDAT- 2013/03/28 06:00
MHDA- 2013/03/28 06:01
PMCR- 2013/03/01
CRDT- 2013/03/28 06:00
PHST- 2013/03/28 06:00 [entrez]
PHST- 2013/03/28 06:00 [pubmed]
PHST- 2013/03/28 06:01 [medline]
PHST- 2013/03/01 00:00 [pmc-release]
AID - S2162-2531(16)30135-4 [pii]
AID - 10.1038/mtna.2013.10 [doi]
PST - epublish
SO  - Mol Ther Nucleic Acids. 2013 Mar 26;2(3):e82. doi: 10.1038/mtna.2013.10.