DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging
In this study, we used bioluminescence imaging (BLI) to track long-term transgene activity following the transfection of brain cells using a nonviral gene therapy technique. Formulations of deoxyribonucleic acid (DNA) combined with 30-mer lysine polymers (substituted with 10 kDa polyethylene glycol)...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
SAGE Publishing
2011-09-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.2310/7290.2010.00053 |
| _version_ | 1826910928619175936 |
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| author | David M. Yurek Anita M. Fletcher Matthew McShane Tomasz H. Kowalczyk Linas Padegimas Marcy R. Weatherspoon Michael D. Kaytor Mark J. Cooper Assem G. Ziady |
| author_facet | David M. Yurek Anita M. Fletcher Matthew McShane Tomasz H. Kowalczyk Linas Padegimas Marcy R. Weatherspoon Michael D. Kaytor Mark J. Cooper Assem G. Ziady |
| author_sort | David M. Yurek |
| collection | DOAJ |
| description | In this study, we used bioluminescence imaging (BLI) to track long-term transgene activity following the transfection of brain cells using a nonviral gene therapy technique. Formulations of deoxyribonucleic acid (DNA) combined with 30-mer lysine polymers (substituted with 10 kDa polyethylene glycol) form nanoparticles that transfect brain cells in vivo and produce transgene activity. Here we show that a single intracerebral injection of these DNA nanoparticles (DNPs) into the rat cortex, striatum, or substantia nigra results in long-term and persistent luciferase transgene activity over an 8- to 11-week period as evaluated by in vivo BLI analysis, and single injections of DNPs into the mouse striatum showed stable luciferase transgene activity for 1 year. Compacted DNPs produced in vivo signals 7- to 34-fold higher than DNA alone. In contrast, ex vivo BLI analysis, which is subject to less signal quenching from surrounding tissues, demonstrated a DNP to DNA alone ratio of 76- to 280-fold. Moreover, the ex vivo BLI analysis confirmed that signals originated from the targeted brain structures. In summary, BLI permits serial analysis of luciferase transgene activity at multiple brain locations following gene transfer with DNPs. Ex vivo analysis may permit more accurate determination of relative activities of gene transfer vectors. |
| first_indexed | 2024-03-07T17:34:10Z |
| format | Article |
| id | doaj.art-74fd1eaad961438ba622f377fd517651 |
| institution | Directory Open Access Journal |
| issn | 1536-0121 |
| language | English |
| last_indexed | 2025-02-17T10:07:10Z |
| publishDate | 2011-09-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj.art-74fd1eaad961438ba622f377fd5176512025-01-02T02:58:12ZengSAGE PublishingMolecular Imaging1536-01212011-09-011010.2310/7290.2010.0005310.2310_7290.2010.00053DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence ImagingDavid M. YurekAnita M. FletcherMatthew McShaneTomasz H. KowalczykLinas PadegimasMarcy R. WeatherspoonMichael D. KaytorMark J. CooperAssem G. ZiadyIn this study, we used bioluminescence imaging (BLI) to track long-term transgene activity following the transfection of brain cells using a nonviral gene therapy technique. Formulations of deoxyribonucleic acid (DNA) combined with 30-mer lysine polymers (substituted with 10 kDa polyethylene glycol) form nanoparticles that transfect brain cells in vivo and produce transgene activity. Here we show that a single intracerebral injection of these DNA nanoparticles (DNPs) into the rat cortex, striatum, or substantia nigra results in long-term and persistent luciferase transgene activity over an 8- to 11-week period as evaluated by in vivo BLI analysis, and single injections of DNPs into the mouse striatum showed stable luciferase transgene activity for 1 year. Compacted DNPs produced in vivo signals 7- to 34-fold higher than DNA alone. In contrast, ex vivo BLI analysis, which is subject to less signal quenching from surrounding tissues, demonstrated a DNP to DNA alone ratio of 76- to 280-fold. Moreover, the ex vivo BLI analysis confirmed that signals originated from the targeted brain structures. In summary, BLI permits serial analysis of luciferase transgene activity at multiple brain locations following gene transfer with DNPs. Ex vivo analysis may permit more accurate determination of relative activities of gene transfer vectors.https://doi.org/10.2310/7290.2010.00053 |
| spellingShingle | David M. Yurek Anita M. Fletcher Matthew McShane Tomasz H. Kowalczyk Linas Padegimas Marcy R. Weatherspoon Michael D. Kaytor Mark J. Cooper Assem G. Ziady DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging Molecular Imaging |
| title | DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging |
| title_full | DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging |
| title_fullStr | DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging |
| title_full_unstemmed | DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging |
| title_short | DNA Nanoparticles: Detection of Long-Term Transgene Activity in Brain using Bioluminescence Imaging |
| title_sort | dna nanoparticles detection of long term transgene activity in brain using bioluminescence imaging |
| url | https://doi.org/10.2310/7290.2010.00053 |
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