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Updated: May 15, 2025

Substrate Generation for Endonucleases of CRISPR/Cas Systems
Published on: September 8, 2012
Click Conjugates of Artificial Ribonucleases: Sequence Specific Cleavage with Multiple Turnover
Sandra Weber1, Timo Weinrich1, Ute Scheffer1
1Institute for Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Max-von-Laue-Strasse 7, D-60438, Frankfurt am Main, Germany.
None:
The azido modified RNA cleaving catalyst 2 has been attached to oligonucleotides containing alkyne linkers in a central position. The resulting conjugates hybridize specifically with complementary RNA strands and cleave them with multiple substrate turnover. RNA half-lives are in the range of 6-7 hours (pH 8, 37 °C). Some well-placed locked nucleic acid (LNA) nucleotides can further increase substrate affinities and reaction rates significantly (t½ of 3.5 hours, kobs = 0.20 h-1). RNA cleavage does not require metal ions and runs equally well in the presence of EDTA. Fragments of precisely defined lengths are formed, well suited for subsequent analysis by mass spectrometry and related bioanalytical techniques.
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