Abstract
Background: The potential for treatment of chronic disorders affecting the CNS is complicated by the inability of several drugs to cross the blood-brain barrier (BBB). None-viral gene therapy applied to brain capillary endothelial cells (BCECs) denotes a novel approach to overcome the restraints in this passage, as turning BCECs into recombinant protein factories by transfection could result in protein secretion into the brain.
Aim: The aim of the present study was to investigate the possibility of transfection to primary rat brain capillary endothelial cells (RBEC) for recombinant protein synthesis.
Results: mRNA expression of proteins with neuroprotective potential in RBEC were enabled. Their expression patters were compared with those of RBE4 and HeLa cells using RT-qPCR analyzes. The evidence for protein synthesis and secretion was obtained by detection of FLAG-tagged to the C-terminal of any of the proteins. Morphological examination of the protein expression was determined using immunofluorescence detecting FLAG. Additionally, the transfection efficiency were determined by Flow cytometry.
Perspective: Our study opens for knowledge on how non-viral gene therapy to BCECs can lead to protein secretion with the perspective of enabling therapeutic protein to target neurons inside the CNS otherwise inhibited to access due to the restraints of the BBB. This therapeutic strategy could be beneficial in treatment of inherited diseases and classical neurodegenerative disorders.
Aim: The aim of the present study was to investigate the possibility of transfection to primary rat brain capillary endothelial cells (RBEC) for recombinant protein synthesis.
Results: mRNA expression of proteins with neuroprotective potential in RBEC were enabled. Their expression patters were compared with those of RBE4 and HeLa cells using RT-qPCR analyzes. The evidence for protein synthesis and secretion was obtained by detection of FLAG-tagged to the C-terminal of any of the proteins. Morphological examination of the protein expression was determined using immunofluorescence detecting FLAG. Additionally, the transfection efficiency were determined by Flow cytometry.
Perspective: Our study opens for knowledge on how non-viral gene therapy to BCECs can lead to protein secretion with the perspective of enabling therapeutic protein to target neurons inside the CNS otherwise inhibited to access due to the restraints of the BBB. This therapeutic strategy could be beneficial in treatment of inherited diseases and classical neurodegenerative disorders.
| Originalsprog | Dansk |
|---|---|
| Publikationsdato | 2014 |
| Status | Udgivet - 2014 |
| Begivenhed | Barriers of the CNS -: Expanding the Understanding of CNS Barriers in Health and Disease - New London, NH, USA Varighed: 15 jun. 2014 → 20 jun. 2014 |
Konference
| Konference | Barriers of the CNS - |
|---|---|
| Land/Område | USA |
| By | New London, NH |
| Periode | 15/06/2014 → 20/06/2014 |