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PMID: 16410612 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Insights into the kinetics of siRNA-mediated gene silencing from live-cell and live-animal bioluminescent imaging.

Nucleic acids research ·Vol. 34 ·No. 1 ·2006-00-00 ·Pages 322-33

Bartlett DW, Davis ME

Abstract

Small interfering RNA (siRNA) molecules are potent effectors of post-transcriptional gene silencing. Using noninvasive bioluminescent imaging and a mathematical model of siRNA delivery and function, the effects of target-specific and treatment-specific parameters on siRNA-mediated gene silencing are monitored in cells stably expressing the firefly luciferase protein. In vitro, luciferase protein levels recover to pre-treatment values within <1 week in rapidly dividing cell lines, but take longer than 3 weeks to return to steady-state levels in nondividing fibroblasts. Similar results are observed in vivo, with knockdown lasting approximately 10 days in subcutaneous tumors in A/J mice and 3-4 weeks in the nondividing hepatocytes of BALB/c mice. These data indicate that dilution due to cell division, and not intracellular siRNA half-life, governs the duration of gene silencing under these conditions. To demonstrate the practical use of the model in treatment design, model calculations are used to predict the dosing schedule required to maintain persistent silencing of target proteins with different half-lives in rapidly dividing or nondividing cells. The approach of bioluminescent imaging combined with mathematical modeling provides useful insights into siRNA function and may help expedite the translation of siRNA into clinically relevant therapeutics for disease treatment and management.

MeSH Terms
Animals Cell Line Cell Proliferation Hepatocytes/metabolism Kinetics Luciferases, Firefly/analysis,genetics Luminescent Agents Mice Mice, Inbred BALB C Models, Biological Neoplasms/genetics,metabolism RNA Interference RNA, Small Interfering/administration & dosage,chemistry
Chemicals
Luminescent Agents RNA, Small Interfering Luciferases, Firefly
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bartlett Derek W
Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Davis Mark E
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkj439
Published
2006-00-00
Epub
2006-00-12
Pages
322-33
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1331994
Subset
IM
Grants
NIBIB NIH HHS · R01 EB004657 · United States
NIBIB NIH HHS · 1 R01 EB004657-01 · United States
External Links
PubMed source DOI: 10.1093/nar/gkj439 PMC Full Text View journal details
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