Nucleic Acids Research, 1993, Vol. 21, No. 6 1381-1387
© 1993
MOLECULAR BIOLOGY |
In vitro selection of fast-hybridizing and effective antisense RNAs directed against the human immunodeficiency virus type 1
Forschungsschwerpunkt Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum Im Neuenheimer Feld 242, D-6900 Heidelberg, Germany 1Max-Planck-Institut für Medizinische Forschung, Abteilung Biophysik, Jahnstraße 29, D-6900 Heidelberg, Germany
*To whom correspondence should be addressed
Received January 4, 1993. Revised February 12, 1993. Accepted February 12, 1993.
The rate of double strand formation between procaryotlc antlsense RNA and complementary RNA In vitro Is known to correlate with the effectiveness of antlsense RNA In vivo. In this work, an In vitro assay for determining the hybridization rates of a large number of antisense RNA species was developed. A set of HIV-1-directed antlsense RNAs with the same 5'-end but successively shortened 3'-ends was produced by alkaline hydrolysis of a 150 nt HIV-1-directed antisense transcript. This mixture was used to determine hybridization rates for individual chain lengths with a complementary HI V-1-derived RNA In vitro. The second order binding rate constants of individual antlsense RNA species differed by more than a factor of 100, although In some cases, slow-hybrldlzlng and fast-hybridizing antlsense RNAs differed by only two or three 3'-termlnally-located nucleotides. The results Indicated that there was not a trivial dependence of binding rates on the chain length of antisense RNAs. Further, the binding rate constants determined In vitro for individual antisense RNA species correlated with the extent of Inhibition of HIV-1 replication In vivo.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  W. James Aptamers in the virologists' toolkit J. Gen. Virol., February 1, 2007; 88(2): 351 - 364. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Kretschmer-Kazemi Far and G. Sczakiel The activity of siRNA in mammalian cells is related to structural target accessibility: a comparison with antisense oligonucleotides Nucleic Acids Res., August 1, 2003; 31(15): 4417 - 4424. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. B. Tummala, N. E. Welker, and E. T. Papoutsakis Design of Antisense RNA Constructs for Downregulation of the Acetone Formation Pathway of Clostridium acetobutylicum J. Bacteriol., March 15, 2003; 185(6): 1923 - 1934. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. H. Lloyd, R. V. Giles, D. G. Spiller, J. Grzybowski, D. M. Tidd, and D. R. Sibson Determination of optimal sites of antisense oligonucleotide cleavage within TNF{alpha} mRNA Nucleic Acids Res., September 1, 2001; 29(17): 3664 - 3673. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. D. Blair, Z. N. Adelman, and K. E. Olson Molecular Strategies for Interrupting Arthropod-Borne Virus Transmission by Mosquitoes Clin. Microbiol. Rev., October 1, 2000; 13(4): 651 - 661. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Patzel and G. Sczakiel In vitro selection supports the view of a kinetic control of antisense RNA-mediated inhibition of gene expression in mammalian cells Nucleic Acids Res., July 1, 2000; 28(13): 2462 - 2466. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Gewirtz, D. L. Sokol, and M. Z. Ratajczak Nucleic Acid Therapeutics: State of the Art and Future Prospects Blood, August 1, 1998; 92(3): 712 - 736. [Full Text] [PDF]
|
|
|
|
|
|
R. Kronenwett, U. Steidl, M. Kirsch, G. Sczakiel, and R. Haas Oligodeoxyribonucleotide Uptake in Primary Human Hematopoietic Cells Is Enhanced by Cationic Lipids and Depends on the Hematopoietic Cell Subset Blood, February 1, 1998; 91(3): 852 - 862. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. F. Lima, V. Brown-Driver, M. Fox, R. Hanecak, and T. W. Bruice Combinatorial Screening and Rational Optimization for Hybridization to Folded Hepatitis C Virus RNA of Oligonucleotides with Biological Antisense Activity J. Biol. Chem., January 3, 1997; 272(1): 626 - 638. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Stein and Y. Cheng Antisense oligonucleotides as therapeutic agents--is the bullet really magical? Science, August 20, 1993; 261(5124): 1004 - 1012. [Abstract] [PDF]
|
|