Site-Specific Cleavage of RNAs Derived from the <i>PIM1</i> 3′-UTR by a Metal-Free Artificial Ribonuclease

Site-Specific Cleavage of RNAs Derived from the <i>PIM1</i> 3′-UTR by a Metal-Free Artificial Ribonuclease

Felix Zellmann, Laura Thomas, Ute Scheffer, Roland K. Hartmann, Michael W. Göbel

Oligonucleotide conjugates of tris(2-aminobenzimidazole) have been reported previously to cleave complementary RNA strands with high levels of sequence and site specificity. The RNA substrates used in these studies were oligonucleotides not longer than 29-mers. Here we show that ~150&#8315;400-mer model transcripts derived from the 3&#8242;-untranslated region of the PIM1 mRNA reacted with rates and specificities comparable to those of short oligonucleotide substrates. The replacement of DNA by DNA/LNA mixmers further increased the cleavage rate. Tris(2-aminobenzimidazoles) were designed to interact with phosphates and phosphate esters. A cell, however, contains large amounts of phosphorylated species that may cause competitive inhibition of RNA cleavage. It is thus important to note that no loss in reaction rates was observed in phosphate buffer. This opens the way to in-cell applications for this type of artificial nuclease. Furthermore, we disclose a new synthetic method giving access to tris(2-aminobenzimidazoles) in multigram amounts.

2019

2-aminobenzimidazole, cleavage of large RNA molecules, cleavage site selection, DNA/LNA mixmers, dye labeling, guanidine analogs, oligonucleotides, specificity of cleavage

DOI: 10.3390/molecules24040807

Molecules

MDPI AG

Organic chemistry

EN