Catalytic RNAs with aminoacylation activity
AssigneeThe Research Foundation of State University of New York
InventorsHiroaki Suga, Dimitrios Kourouklis, Hirohide Saito, Nick Lee, Neil Bonzagni
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Abstract
The present invention provides catalytic RNA molecules having cis or trans aminoacylation activity. The catalytic RNA molecules having cis aminoacylation activity comprise a catalytic domain and an aminoacylation domain. The catalytic RNA molecules having trans aminoacylation activity only have the catalytic domain. A method is provided for constructing and screening of these molecules. These molecules are suitable for aminoacylating tRNA-like molecules with specific amino acids.
Claims9
A method for identifying cis-aminoacylating catalytic RNA molecules comprising the steps of:
- a. providing tRNA molecules;
- b. providing ribozyme molecules;
- c. attaching the ribozyme molecules to the 5′ end of the tRNA molecules to obtain a pool of ribozyme-tRNA molecules;
- d. contacting the ribozyme-tRNA molecules with an amino acid substrate wherein the amino acid substrate is selected from the group consisting of biotinyl-aminoacyl-cyanomethyl esters, aminoacyl-thio esters and aminoacyl-adenylates;
- and
- e. partitioning aminoacylated RNA molecules from non-aminoacylated RNA molecules wherein the aminoacylated RNA molecules are cis-aminoacylating catalytic RNA molecules aminoacylated with the amino acid substrate at the 3′ end of the ribozyme-tRNA molecules.
A method for identifying cis-aminoacylating catalytic RNA molecules comprising the steps of:
- a. providing RNA molecules having a tRNA domain and a 5′-leader ribozyme;
- b. contacting the RNA molecules with an amino acid substrate wherein the amino acid substrate is not conjugated to RNA selected from the group consisting of biotinyl-aminoacyl cyanomethyl esters, aminoacyl-thio esters and aminoacyl-adenylates;
- and
- c. partitioning aminoacylated RNA molecules from non-aminoacylated RNA molecules wherein the aminoacylated RNA molecules are cis-aminoacylating catalytic RNA molecules aminoacylated with the amino acid substrate at the 3′ end of the ribozyme-tRNA molecules.
Dependencies are inferred from the claim text.