Use of genetically induced repression of ribonucleotide reductase as target in trypanocide formulation

Sulaiman Faoziyat A.¹*, Akanji M.A.¹, Green, B. O.², Nguyen B.³, Nguyen T.³ and Gunzl A.³

¹Biochemistry Department, University of Ilorin, PMB 1515 Ilorin, Nigeria.

²College of Naturopathic Medicine, University of Bridgeport, Connecticut, U.S.A.

³Genetics and Developmental Biology, University of Connecticut, Farmington USA.

*Corresponding author E-mail: faoziyat20022002@yahoo.com

Received 22 February, 2012; Accepted 02 March, 2012

Abstract

In an in vitro experiment, ribonucleotide reductase was genetically validated as a chemotherapeutic target by engineering a plasmid vector (Recombinant DNA) for conditional expression silencing of the enzyme, by cloning the ribonucleotide reductase in a polymerase chain reaction (PCR). It was then amplified, purified and inserted into the plasmids to yield the recombinant DNA of interest, which was transfected into the parasites (clonal transfected cell lines) by electroporation. Parasite cell growth was monitored upon RNAi (Ribonucleotide reductase Interference) mediated silencing of RNR expression. The results generated showed that mRNA repression in the four groups were time dependent in parasites induced at 24hrs, 42 and 48 hours, with 48 hours cDNA showing the most significant mRNA repression. Ribonucleotide reductase was concluded to be an essential enzyme in the trypanosomes as depicted by the RNAi mediated silencing of its expression. It is also a very good drug target.

Keywords: Ribonucleotide reductase, mRNA, cDNA, Recombinant DNA, RNAi, PCR.