Published online 12 January 2006
Article |
Insights into the kinetics of siRNA-mediated gene silencing from live-cell and live-animal bioluminescent imaging
Chemical Engineering, California Institute of Technology Pasadena, CA 91125, USA
*To whom correspondence should be addressed. Tel: +1 626 395 4251; Fax: +1 626 568 8743; Email: mdavis{at}cheme.caltech.edu
Received December 2, 2005. Revised December 23, 2005. Accepted December 23, 2005.
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 
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.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Lasham, M. Herbert, N. Coppieters 't Wallant, R. Patel, S. Feng, M. Eszes, H. Cao, and G. Reid A rapid and sensitive method to detect siRNA-mediated mRNA cleavage in vivo using 5' RACE and a molecular beacon probe Nucleic Acids Res., November 26, 2009; (2009) gkp1076v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Hashimoto, J. Igarashi, and H. Kosaka Sphingosine kinase is induced in mouse 3T3-L1 cells and promotes adipogenesis J. Lipid Res., April 1, 2009; 50(4): 602 - 610. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Behl, P. Krothapalli, T. Desta, S. Roy, and D. T. Graves FOXO1 Plays an Important Role in Enhanced Microvascular Cell Apoptosis and Microvascular Cell Loss in Type 1 and Type 2 Diabetic Rats Diabetes, April 1, 2009; 58(4): 917 - 925. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. S. Bathula, S. H. Garrett, X. D. Zhou, M. A. Sens, D. A. Sens, and S. Somji Cadmium, Vectorial Active Transport, and MT-3-Dependent Regulation of Cadherin Expression in Human Proximal Tubular Cells Toxicol. Sci., April 1, 2008; 102(2): 310 - 318. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. F. Pirollo and E. H. Chang Targeted Delivery of Small Interfering RNA: Approaching Effective Cancer Therapies Cancer Res., March 1, 2008; 68(5): 1247 - 1250. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. J. Schmidt, M. Romanov, O. Ryder, V. Magrini, M. Hickenbotham, J. Glasscock, S. McGrath, E. Mardis, and L. D. Stein Gallus GBrowse: a unified genomic database for the chicken Nucleic Acids Res., January 11, 2008; 36(suppl_1): D719 - D723. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Zhu, J. D. Bjorge, and D. J. Fujita PTP1B Contributes to the Oncogenic Properties of Colon Cancer Cells through Src Activation Cancer Res., November 1, 2007; 67(21): 10129 - 10137. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Bhindi, R. G. Fahmy, H. C. Lowe, C. N. Chesterman, C. R. Dass, M. J. Cairns, E. G. Saravolac, L.-Q. Sun, and L. M. Khachigian Brothers in Arms: DNA Enzymes, Short Interfering RNA, and the Emerging Wave of Small-Molecule Nucleic Acid-Based Gene-Silencing Strategies Am. J. Pathol., October 1, 2007; 171(4): 1079 - 1088. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. W. Bartlett, H. Su, I. J. Hildebrandt, W. A. Weber, and M. E. Davis Impact of tumor-specific targeting on the biodistribution and efficacy of siRNA nanoparticles measured by multimodality in vivo imaging PNAS, September 25, 2007; 104(39): 15549 - 15554. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Sundaram, L. K. Lee, and C. M. Roth Interplay of polyethyleneimine molecular weight and oligonucleotide backbone chemistry in the dynamics of antisense activity Nucleic Acids Res., July 26, 2007; 35(13): 4396 - 4408. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Lin and U. W. Goodenough Gametogenesis in the Chlamydomonas reinhardtii minus Mating Type Is Controlled by Two Genes, MID and MTD1 Genetics, June 1, 2007; 176(2): 913 - 925. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Heidel, Z. Yu, J. Y.-C. Liu, S. M. Rele, Y. Liang, R. K. Zeidan, D. J. Kornbrust, and M. E. Davis Inaugural Article: Administration in non-human primates of escalating intravenous doses of targeted nanoparticles containing ribonucleotide reductase subunit M2 siRNA PNAS, April 3, 2007; 104(14): 5715 - 5721. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Sharma, J. Masri, O. D. Jo, A. Bernath, J. Martin, A. Funk, and J. Gera Protein Kinase C Regulates Internal Initiation of Translation of the GATA-4 mRNA following Vasopressin-induced Hypertrophy of Cardiac Myocytes J. Biol. Chem., March 30, 2007; 282(13): 9505 - 9516. [Abstract] [Full Text] [PDF]
|
|