RNase P, a ribonucleoprotein responsible for the 5´ maturation of precursor tRNAs (ptRNAs) in all organisms, can be enticed to cleave any target mRNA that forms a ptRNA-like structure and sequence-specific complex when bound to an RNA, termed the EGS (external guide sequence). In the present study, F3H (flavanone 3-hydroxylase), a key enzyme in the flavonoid biosynthetic pathway that participates in the formation of red-coloured anthocyanins, was used as a target for RNase P-mediated gene disruption in maize cells. Transient expression of an EGS complementary to the F3H mRNA resulted in suppression of F3H to 29% of the control, as indicated by a reduced number of anthocyanin-accumulating cells. This decrease was not observed in experiments where a disabled mutant EGS was expressed. Our results demonstrate the potential of employing plant RNase P, in the presence of an appropriate gene-specific EGS, as a tool for targeted degradation of mRNAs.
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June 15 2004
RNase P as a tool for disruption of gene expression in maize cells Available to Purchase
Sunita RANGARAJAN;
Sunita RANGARAJAN
*Department of Biochemistry, The Ohio State University, 707 Biological Sciences Building, 484 West 12th Avenue, Columbus, OH 43210, U.S.A.
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M. L. Stephen RAJ;
M. L. Stephen RAJ
*Department of Biochemistry, The Ohio State University, 707 Biological Sciences Building, 484 West 12th Avenue, Columbus, OH 43210, U.S.A.
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J. Marcela HERNANDEZ;
J. Marcela HERNANDEZ
†Department of Plant Cellular and Molecular Biology, and Plant Biotechnology Center, 206 Rightmire Hall, 1060 Carmack Road, The Ohio State University, Columbus, OH 43210, U.S.A.
‡The Ohio State Biochemistry Program, 108 Aronoff Building, 318 West 12th Avenue, The Ohio State University, Columbus, OH 43210, U.S.A.
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Erich GROTEWOLD;
Erich GROTEWOLD
†Department of Plant Cellular and Molecular Biology, and Plant Biotechnology Center, 206 Rightmire Hall, 1060 Carmack Road, The Ohio State University, Columbus, OH 43210, U.S.A.
‡The Ohio State Biochemistry Program, 108 Aronoff Building, 318 West 12th Avenue, The Ohio State University, Columbus, OH 43210, U.S.A.
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Venkat GOPALAN
Venkat GOPALAN
1
*Department of Biochemistry, The Ohio State University, 707 Biological Sciences Building, 484 West 12th Avenue, Columbus, OH 43210, U.S.A.
†Department of Plant Cellular and Molecular Biology, and Plant Biotechnology Center, 206 Rightmire Hall, 1060 Carmack Road, The Ohio State University, Columbus, OH 43210, U.S.A.
‡The Ohio State Biochemistry Program, 108 Aronoff Building, 318 West 12th Avenue, The Ohio State University, Columbus, OH 43210, U.S.A.
1To whom correspondence should be addressed (email [email protected]).
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Publisher: Portland Press Ltd
Received:
March 18 2004
Revision Received:
April 15 2004
Accepted:
April 20 2004
Accepted Manuscript online:
April 20 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2004
2004
Biochem J (2004) 380 (3): 611–616.
Article history
Received:
March 18 2004
Revision Received:
April 15 2004
Accepted:
April 20 2004
Accepted Manuscript online:
April 20 2004
Citation
Sunita RANGARAJAN, M. L. Stephen RAJ, J. Marcela HERNANDEZ, Erich GROTEWOLD, Venkat GOPALAN; RNase P as a tool for disruption of gene expression in maize cells. Biochem J 15 June 2004; 380 (3): 611–616. doi: https://doi.org/10.1042/bj20040442
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