Whole animal in vivo imaging after transient, nonviral gene delivery to the rat central nervous system

Ellen S. Hauck, Shaomin Zou, Keith Scarfo, Michael H. Nantz, James G. Hecker

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

We previously showed that a vector:lipid delivery system, comprised of a plasmid DNA vector and cationic lipid (lipoplex), when injected into the cerebrospinal fluid (CSF) of rats can deliver reporter genes in vivo efficiently and with widespread expression to the Central Nervous System (CNS). To further characterize this delivery system, we now present experiments that demonstrate the in vivo time-to-peak expression of the reporter gene, firefly luciferase. We infused a formulated lipoplex containing the lipid MLRI [dissymmetric myristoyl (14:0) and lauroyl (12:1) rosenthal inhibitor-substituted compound formed from the tetraalkylammonium glycerol-based DORI] and pNDluc, a luciferase vector, into CSF in the cisterna magna (CM) of the rat. Luciferase activity was followed over time by bioluminescence imaging after injection of luciferin. Our results show that luciferase activity in the CNS of rats is widespread, peaks 72 hours after injection into CM and can be detected in vivo for at least 7-10 days after peak expression. We further show that in contrast to injection into CSF, enzyme activity is not widely distributed after injection of the vector into brain parenchyma, emphasizing the importance of CSF delivery to achieve widespread vector distribution. Finally, we confirm the distribution of firefly luciferase in brain by immunohistochemical staining from an animal that was euthanized at the peak of enzyme expression.

Original languageEnglish (US)
Pages (from-to)1857-1864
Number of pages8
JournalMolecular Therapy
Volume16
Issue number11
DOIs
StatePublished - 2008
Externally publishedYes

Fingerprint

Cerebrospinal Fluid
Central Nervous System
Luciferases
Cisterna Magna
Firefly Luciferases
Injections
Lipids
Reporter Genes
Genes
Brain
Enzymes
Glycerol
Plasmids
Staining and Labeling
DNA

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Genetics
  • Drug Discovery
  • Pharmacology

Cite this

Whole animal in vivo imaging after transient, nonviral gene delivery to the rat central nervous system. / Hauck, Ellen S.; Zou, Shaomin; Scarfo, Keith; Nantz, Michael H.; Hecker, James G.

In: Molecular Therapy, Vol. 16, No. 11, 2008, p. 1857-1864.

Research output: Contribution to journalArticle

Hauck, Ellen S. ; Zou, Shaomin ; Scarfo, Keith ; Nantz, Michael H. ; Hecker, James G. / Whole animal in vivo imaging after transient, nonviral gene delivery to the rat central nervous system. In: Molecular Therapy. 2008 ; Vol. 16, No. 11. pp. 1857-1864.
@article{73a411a1515c4768bea832c185b64301,
title = "Whole animal in vivo imaging after transient, nonviral gene delivery to the rat central nervous system",
abstract = "We previously showed that a vector:lipid delivery system, comprised of a plasmid DNA vector and cationic lipid (lipoplex), when injected into the cerebrospinal fluid (CSF) of rats can deliver reporter genes in vivo efficiently and with widespread expression to the Central Nervous System (CNS). To further characterize this delivery system, we now present experiments that demonstrate the in vivo time-to-peak expression of the reporter gene, firefly luciferase. We infused a formulated lipoplex containing the lipid MLRI [dissymmetric myristoyl (14:0) and lauroyl (12:1) rosenthal inhibitor-substituted compound formed from the tetraalkylammonium glycerol-based DORI] and pNDluc, a luciferase vector, into CSF in the cisterna magna (CM) of the rat. Luciferase activity was followed over time by bioluminescence imaging after injection of luciferin. Our results show that luciferase activity in the CNS of rats is widespread, peaks 72 hours after injection into CM and can be detected in vivo for at least 7-10 days after peak expression. We further show that in contrast to injection into CSF, enzyme activity is not widely distributed after injection of the vector into brain parenchyma, emphasizing the importance of CSF delivery to achieve widespread vector distribution. Finally, we confirm the distribution of firefly luciferase in brain by immunohistochemical staining from an animal that was euthanized at the peak of enzyme expression.",
author = "Hauck, {Ellen S.} and Shaomin Zou and Keith Scarfo and Nantz, {Michael H.} and Hecker, {James G.}",
year = "2008",
doi = "10.1038/mt.2008.183",
language = "English (US)",
volume = "16",
pages = "1857--1864",
journal = "Molecular Therapy",
issn = "1525-0016",
publisher = "Nature Publishing Group",
number = "11",

}

TY - JOUR

T1 - Whole animal in vivo imaging after transient, nonviral gene delivery to the rat central nervous system

AU - Hauck, Ellen S.

AU - Zou, Shaomin

AU - Scarfo, Keith

AU - Nantz, Michael H.

AU - Hecker, James G.

PY - 2008

Y1 - 2008

N2 - We previously showed that a vector:lipid delivery system, comprised of a plasmid DNA vector and cationic lipid (lipoplex), when injected into the cerebrospinal fluid (CSF) of rats can deliver reporter genes in vivo efficiently and with widespread expression to the Central Nervous System (CNS). To further characterize this delivery system, we now present experiments that demonstrate the in vivo time-to-peak expression of the reporter gene, firefly luciferase. We infused a formulated lipoplex containing the lipid MLRI [dissymmetric myristoyl (14:0) and lauroyl (12:1) rosenthal inhibitor-substituted compound formed from the tetraalkylammonium glycerol-based DORI] and pNDluc, a luciferase vector, into CSF in the cisterna magna (CM) of the rat. Luciferase activity was followed over time by bioluminescence imaging after injection of luciferin. Our results show that luciferase activity in the CNS of rats is widespread, peaks 72 hours after injection into CM and can be detected in vivo for at least 7-10 days after peak expression. We further show that in contrast to injection into CSF, enzyme activity is not widely distributed after injection of the vector into brain parenchyma, emphasizing the importance of CSF delivery to achieve widespread vector distribution. Finally, we confirm the distribution of firefly luciferase in brain by immunohistochemical staining from an animal that was euthanized at the peak of enzyme expression.

AB - We previously showed that a vector:lipid delivery system, comprised of a plasmid DNA vector and cationic lipid (lipoplex), when injected into the cerebrospinal fluid (CSF) of rats can deliver reporter genes in vivo efficiently and with widespread expression to the Central Nervous System (CNS). To further characterize this delivery system, we now present experiments that demonstrate the in vivo time-to-peak expression of the reporter gene, firefly luciferase. We infused a formulated lipoplex containing the lipid MLRI [dissymmetric myristoyl (14:0) and lauroyl (12:1) rosenthal inhibitor-substituted compound formed from the tetraalkylammonium glycerol-based DORI] and pNDluc, a luciferase vector, into CSF in the cisterna magna (CM) of the rat. Luciferase activity was followed over time by bioluminescence imaging after injection of luciferin. Our results show that luciferase activity in the CNS of rats is widespread, peaks 72 hours after injection into CM and can be detected in vivo for at least 7-10 days after peak expression. We further show that in contrast to injection into CSF, enzyme activity is not widely distributed after injection of the vector into brain parenchyma, emphasizing the importance of CSF delivery to achieve widespread vector distribution. Finally, we confirm the distribution of firefly luciferase in brain by immunohistochemical staining from an animal that was euthanized at the peak of enzyme expression.

UR - http://www.scopus.com/inward/record.url?scp=54949094662&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=54949094662&partnerID=8YFLogxK

U2 - 10.1038/mt.2008.183

DO - 10.1038/mt.2008.183

M3 - Article

C2 - 18728638

AN - SCOPUS:54949094662

VL - 16

SP - 1857

EP - 1864

JO - Molecular Therapy

JF - Molecular Therapy

SN - 1525-0016

IS - 11

ER -