Highly efficient transfection of rat cortical neurons using carbosilane dendrimers unveils a neuroprotective role for HIF-1alpha in early chemical hypoxia-mediated neurotoxicity.
OBJECTIVE
To study the effect of a non-viral vector (carbosilane dendrimer) to efficiently deliver small interfering RNA to postmitotic neurons to study the function of hypoxia-inducible factor-1alpha (HIF1-alpha) during chemical hypoxia-mediated neurotoxicity.


METHODS
Chemical hypoxia was induced in primary rat cortical neurons by exposure to CoCl(2). HIF1-alpha levels were determined by Western Blot and toxicity was evaluated by both MTT and LDH assays. Neurons were incubated with dendriplexes containing anti-HIF1-alpha siRNA and both uptake and HIF1-alpha knockdown efficiency were evaluated.


RESULTS
We report that a non-viral vector (carbosilane dendrimer) can deliver specific siRNA to neurons and selectively block HIF1-alpha synthesis with similar efficiency to that achieved by viral vectors. Using this method, we have found that this transcription factor plays a neuroprotective role during the early phase of chemical hypoxia-mediated neurotoxicity.


CONCLUSIONS
This work represents a proof-of-concept for the use of carbosilane dendrimers to deliver specific siRNA to postmitotic neurons to block selected protein synthesis. This indicates that this type of vector is a good alternative to viral vectors to achieve very high transfection levels in neurons. This also suggests that carbosilane dendrimers might be very useful for gene therapy.
Posadas
Inmaculada
I
Departamento de Ciencias Médicas, Unidad Asociada Neurodeath, CSIC-UCLM, Universidad de Castilla-La Mancha, Avda. Almansa, 14, 02006, Albacete, Spain.
López-Hernández
Beatriz
B
Clemente
Maria Isabel
MI
Jiménez
Jose Luis
JL
Ortega
Paula
P
de la Mata
Javier
J
Gómez
Rafael
R
Muñoz-Fernández
María Angeles
MA
Ceña
Valentín
V
D000818:Animals; D015687:Cell Hypoxia / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002540:Cerebral Cortex / Q000166:cytology; D003035:Cobalt; D050091:Dendrimers / Q000737:chemistry / Q000633:toxicity*; D055785:Gene Knockdown Techniques; D015316:Genetic Therapy; D000860:Hypoxia / Q000139:chemically induced / Q000235:genetics* / Q000628:therapy*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000008:administration & dosage / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012821:Silanes / Q000737:chemistry / Q000633:toxicity*; D014162:Transfection
D050091:Dendrimers; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D018696:Neuroprotective Agents; D034741:RNA, Small Interfering; D012821:Silanes; C504072:carbosilane; D003035:Cobalt