Nucleic Acids Research, 2003, Vol. 31, No. 3 e8
© 2003 Oxford University Press
Efficient incorporation of CoA, NAD and FAD into RNA by in vitro transcription
Department of Chemistry and Biochemistry, University of Southern Mississippi, Hattiesburg, MS 39406-5043, USA
*Tel: +1 601 266 4371; Fax: +1 601 266 6075; Email: faqing.h.huang{at}usm.edu
Protein enzymes frequently recruit small molecule coenzymes to perform a variety of biochemical reactions. While the catalytic activities of RNA have been expanding rapidly, a similar strategy for RNA to utilize coenzymes and to increase its functional capabilities has yet to be demonstrated. A general in vitro transcription procedure has been developed to efficiently prepare RNA with coenzymes CoA, NAD and FAD covalently attached to the 5' end. These adenosine-containing coenzymes initiate transcription under the T7 class II promoter by T7 RNA polymerase. In addition to the three coenzymes, other adenosine-containing molecules may be incorporated into the first nucleotide position of RNA as well. This method provides easy access to CoA–, NAD– and FAD–RNA, which may find broad applications in generating coenzyme- utilizing ribozymes. In addition, both oxidized FAD and reduced NADH are highly fluorescent. NADH–RNA and FAD–RNA can therefore be used as probes for DNA/RNA detection and for structural investigation of RNA function by fluorescence spectroscopy.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  K. Onizuka, Y. Taniguchi, and S. Sasaki A new usage of functionalized oligodeoxynucleotide probe for site-specific modification of a guanine base within RNA Nucleic Acids Res., March 1, 2010; 38(5): 1760 - 1766. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Fujita, H. Furuta, and Y. Ikawa Evolutionary optimization of a modular ligase ribozyme: a small catalytic unit and a hairpin motif masking an element that could form an inactive structure Nucleic Acids Res., January 27, 2010; (2010): gkq018v1 - gkq018. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. E. Kowtoniuk, Y. Shen, J. M. Heemstra, I. Agarwal, and D. R. Liu A chemical screen for biological small molecule-RNA conjugates reveals CoA-linked RNA PNAS, May 12, 2009; 106(19): 7768 - 7773. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Fujita, H. Furuta, and Y. Ikawa Tailoring RNA modular units on a common scaffold: A modular ribozyme with a catalytic unit for {beta}-nicotinamide mononucleotide-activated RNA ligation RNA, May 1, 2009; 15(5): 877 - 888. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Zhang, D. Shu, F. Huang, and P. Guo Instrumentation and metrology for single RNA counting in biological complexes or nanoparticles by a single-molecule dual-view system RNA, October 1, 2007; 13(10): 1793 - 1802. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. TORTORICI, B. A. SHAPIRO, and J. T. PATTON A base-specific recognition signal in the 5' consensus sequence of rotavirus plus-strand RNAs promotes replication of the double-stranded RNA genome segments RNA, January 1, 2006; 12(1): 133 - 146. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Li, C. Yu, and F. Huang Novel cyanine-AMP conjugates for efficient 5' RNA fluorescent labeling by one-step transcription and replacement of [{gamma}-32P]ATP in RNA structural investigation Nucleic Acids Res., February 24, 2005; 33(4): e37 - e37. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Coleman, G. Wang, and F. Huang Superior 5' homogeneity of RNA from ATP-initiated transcription under the T7 {phi}2.5 promoter Nucleic Acids Res., January 15, 2004; 32(1): e14 - e14. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. HUANG, G. WANG, T. COLEMAN, and N. LI Synthesis of adenosine derivatives as transcription initiators and preparation of 5' fluorescein- and biotin-labeled RNA through one-step in vitro transcription RNA, December 1, 2003; 9(12): 1562 - 1570. [Abstract] [Full Text] [PDF]
|
|