Diabetes
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Bain, J. R.
Right arrow Articles by Becker, T. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bain, J. R.
Right arrow Articles by Becker, T. C.
Social Bookmarking
Add to CiteULike   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Diabetes 53:2190-2194, 2004
© 2004 by the American Diabetes Association, Inc.

An Adenovirus Vector for Efficient RNA Interference–Mediated Suppression of Target Genes in Insulinoma Cells and Pancreatic Islets of Langerhans

James R. Bain1,2, Jonathan C. Schisler1,3, Koji Takeuchi1,2, Christopher B. Newgard1,2,4, and Thomas C. Becker1,2,4

1 Sarah W. Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, North Carolina
2 Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina
3 University of Texas Southwestern Medical Center at Dallas, Dallas, Texas
4 Division of Endocrinology, Metabolism, and Nutrition, Department of Medicine, Duke University Medical Center, Durham, North Carolina

Silencing gene expression by RNA interference (RNAi) can provide insight into gene function but requires efficient delivery of small interfering RNAs (siRNAs) into cells. Introduction of exogenous nucleic acids can be especially difficult in cultured pancreatic islets. This article describes a method for making recombinant adenoviruses that efficiently drive expression of siRNAs in islet ß-cells and a ß-cell–derived cell line. Transduction with a virus expressing an siRNA specific for GLUT2 reduced GLUT2 mRNA and protein levels by 80% in the INS-1–derived ß-cell line, 832/13, and GLUT2 protein levels by >90% in primary rat islets. Another virus expressing an siRNA specific for glucokinase (GK) caused 80% suppression of GK mRNA and 50% suppression of GK protein levels in 832/13 cells. These experiments validate recombinant adenoviral RNAi vectors as a useful tool for suppression of the expression of specific genes in pancreatic islets and ß-cell lines. Advantages of this approach include 1) the high efficiency of adenovirus-mediated gene transfer in insulinoma cell lines and rat islets and 2) the rapidity with which RNAi constructs can be prepared and tested relative to stable-transfection strategies.

Address correspondence reprint requests to Thomas C. Becker, PhD, Duke Independence Park Facility, Duke University Medical Center, 4321 Medical Park Dr., Suite 200, Durham, NC 27704. E-mail: thomas.becker{at}duke.edu


Add to CiteULike CiteULike   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 DiabetesHome page
B. E. Levin, T. C. Becker, J.-i. Eiki, B. B. Zhang, and A. A. Dunn-Meynell
Ventromedial Hypothalamic Glucokinase Is an Important Mediator of the Counterregulatory Response to Insulin-Induced Hypoglycemia
Diabetes, May 1, 2008; 57(5): 1371 - 1379.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. W. Joseph, M. L. Odegaard, S. M. Ronnebaum, S. C. Burgess, J. Muehlbauer, A. D. Sherry, and C. B. Newgard
Normal Flux through ATP-Citrate Lyase or Fatty Acid Synthase Is Not Required for Glucose-stimulated Insulin Secretion
J. Biol. Chem., October 26, 2007; 282(43): 31592 - 31600.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. M. Schroeder-Gloeckler, S. M. Rahman, R. C. Janssen, L. Qiao, J. Shao, M. Roper, S. J. Fischer, E. Lowe, D. J. Orlicky, J. L. McManaman, et al.
CCAAT/Enhancer-binding Protein beta Deletion Reduces Adiposity, Hepatic Steatosis, and Diabetes in Leprdb/db Mice
J. Biol. Chem., May 25, 2007; 282(21): 15717 - 15729.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
O. T. Hardy, H. E. Hohmeier, T. C. Becker, E. Manduchi, N. M. Doliba, R. K. Gupta, P. White, C. J. Stoeckert Jr, F. M. Matschinsky, C. B. Newgard, et al.
Functional Genomics of the {beta}-Cell: Short-Chain 3-Hydroxyacyl-Coenzyme A Dehydrogenase Regulates Insulin Secretion Independent of K+ Currents
Mol. Endocrinol., March 1, 2007; 21(3): 765 - 773.
[Abstract] [Full Text] [PDF]

Home page
 J Mol EndocrinolHome page
R. Takahashi, H. Ishihara, K. Takahashi, A. Tamura, S. Yamaguchi, T. Yamada, H. Katagiri, and Y. Oka
Efficient and controlled gene expression in mouse pancreatic islets by arterial delivery of tetracycline-inducible adenoviral vectors
J. Mol. Endocrinol., January 1, 2007; 38(1): 127 - 136.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. W. Joseph, M. V. Jensen, O. Ilkayeva, F. Palmieri, C. Alarcon, C. J. Rhodes, and C. B. Newgard
The Mitochondrial Citrate/Isocitrate Carrier Plays a Regulatory Role in Glucose-stimulated Insulin Secretion
J. Biol. Chem., November 24, 2006; 281(47): 35624 - 35632.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. M. Ronnebaum, O. Ilkayeva, S. C. Burgess, J. W. Joseph, D. Lu, R. D. Stevens, T. C. Becker, A. D. Sherry, C. B. Newgard, and M. V. Jensen
A Pyruvate Cycling Pathway Involving Cytosolic NADP-dependent Isocitrate Dehydrogenase Regulates Glucose-stimulated Insulin Secretion
J. Biol. Chem., October 13, 2006; 281(41): 30593 - 30602.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. V. Jensen, J. W. Joseph, O. Ilkayeva, S. Burgess, D. Lu, S. M. Ronnebaum, M. Odegaard, T. C. Becker, A. D. Sherry, and C. B. Newgard
Compensatory Responses to Pyruvate Carboxylase Suppression in Islet beta-Cells: PRESERVATION OF GLUCOSE-STIMULATED INSULIN SECRETION
J. Biol. Chem., August 4, 2006; 281(31): 22342 - 22351.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
J. C. Raum, K. Gerrish, I. Artner, E. Henderson, M. Guo, L. Sussel, J. C. Schisler, C. B. Newgard, and R. Stein
FoxA2, Nkx2.2, and PDX-1 Regulate Islet {beta}-Cell-Specific mafA Expression through Conserved Sequences Located between Base Pairs -8118 and -7750 Upstream from the Transcription Start Site
Mol. Cell. Biol., August 1, 2006; 26(15): 5735 - 5743.
[Abstract] [Full Text] [PDF]

Home page
 Pharmacol. Rev.Home page
A. S. Narang and R. I. Mahato
Biological and biomaterial approaches for improved islet transplantation.
Pharmacol. Rev., June 1, 2006; 58(2): 194 - 243.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Chen, J.-h. Ding, R. Bekeredjian, B.-z. Yang, R. V. Shohet, S. A. Johnston, H. E. Hohmeier, C. B. Newgard, and P. A. Grayburn
Efficient gene delivery to pancreatic islets with ultrasonic microbubble destruction technology
PNAS, May 30, 2006; 103(22): 8469 - 8474.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W. Cao, Q. F. Collins, T. C. Becker, J. Robidoux, E. G. Lupo Jr., Y. Xiong, K. W. Daniel, L. Floering, and S. Collins
p38 Mitogen-activated Protein Kinase Plays a Stimulatory Role in Hepatic Gluconeogenesis
J. Biol. Chem., December 30, 2005; 280(52): 42731 - 42737.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Diao, Z. Asghar, C. B. Chan, and M. B. Wheeler
Glucose-regulated Glucagon Secretion Requires Insulin Receptor Expression in Pancreatic {alpha}-Cells
J. Biol. Chem., September 30, 2005; 280(39): 33487 - 33496.
[Abstract] [Full Text] [PDF]

Home page
 DiabetesHome page
R. Wang, J. Li, K. Lyte, N. K. Yashpal, F. Fellows, and C. G. Goodyer
Role for {beta}1 Integrin and Its Associated {alpha}3, {alpha}5, and {alpha}6 Subunits in Development of the Human Fetal Pancreas
Diabetes, July 1, 2005; 54(7): 2080 - 2089.
[Abstract] [Full Text] [PDF]

Home page
 DiabetesHome page
K. N. Samli, M. J. McGuire, C. B. Newgard, S. A. Johnston, and K. C. Brown
Peptide-Mediated Targeting of the Islets of Langerhans
Diabetes, July 1, 2005; 54(7): 2103 - 2108.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. Iype, J. Francis, J. C. Garmey, J. C. Schisler, R. Nesher, G. C. Weir, T. C. Becker, C. B. Newgard, S. C. Griffen, and R. G. Mirmira
Mechanism of insulin Gene Regulation by the Pancreatic Transcription Factor Pdx-1: APPLICATION OF PRE-mRNA ANALYSIS AND CHROMATIN IMMUNOPRECIPITATION TO ASSESS FORMATION OF FUNCTIONAL TRANSCRIPTIONAL COMPLEXES
J. Biol. Chem., April 29, 2005; 280(17): 16798 - 16807.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
N. K. Yashpal, J. Li, M. B. Wheeler, and R. Wang
Expression of {beta}1 Integrin Receptors during Rat Pancreas Development--Sites and Dynamics
Endocrinology, April 1, 2005; 146(4): 1798 - 1807.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Diabetes Diabetes Care Clinical Diabetes Diabetes Spectrum
Copyright © 2004 by the American Diabetes Association.