Artificial nucleases that selectively cleave HIV-1 Rev Response Element (RRE) mRNA were synthesized.  Chelators EDTA, DTPA, NTA, DOTA, GGH, and KGHK were linked to a Rev-derived peptide; the coordination complexes with Fe2+, Co2+, Ni2+, and Cu2+ were observed to bind RRE RNA with high affinity (KD~0.2�16 nM), with weaker binding to a low-affinity site (KD~1000 nM) and non-targeted DNA (KD~1700�3600 nM).  Second order rate constants of oxidative scission of the RNA up to 460,000 M-1min-1 were observed (Cu-NTA-Rev).  Selectivity of RRE RNA scission was demonstrated by Rev targeting, tRNA competition, and reactivity of RRE RNA vs non-targeted DNA.  Optimal activity was observed for catalysts with reduction potentials between those of the redox coreagents;  ROS produced by the catalysts were responsible for RNA scission.  MADLI-TOF MS revealed H-4 and H-5 abstraction near each metal center, and the distance-dependence of reactivity allowed characterization of active ROS. 

Targeted scission of HIV RRE RNA by Rev-coupled transition metal chelates

244th National Meeting (2012)

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