RNase P as a tool for disruption of gene expression in maize cells.
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.
Rangarajan
Sunita
S
Department of Biochemistry, The Ohio State University, 707 Biological Sciences Building, 484 West 12th Avenue, Columbus, OH 43210, USA.
Raj
M L Stephen
ML
Hernandez
J Marcela
JM
Grotewold
Erich
E
Gopalan
Venkat
V
D018506:Gene Expression Regulation, Plant / Q000187:drug effects*; D017343:Genes, Plant / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation / Q000187:drug effects; D018749:RNA, Plant / Q000096:biosynthesis / Q000378:metabolism; D012107:Research Design; D043262:Ribonuclease P / Q000378:metabolism / Q000494:pharmacology*; D003313:Zea mays / Q000187:drug effects* / Q000201:enzymology
D018749:RNA, Plant; D006899:Mixed Function Oxygenases; C048797:flavanone 3-dioxygenase; D043262:Ribonuclease P