Method for Designing RNA Binding Protein Utilizing PPR Motif, and Use Thereof

摘要

A method for designing a protein capable of binding in an RNA base selective manner or RNA base sequence specific manner is provided. The protein of the present invention is a protein containing one or more of PPR motifs (preferably 2 to 14 PPR motifs) each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1 (wherein Helix A is a moiety of 12-amino acid length capable of forming an α-helix structure, and is represented by the formula 2, wherein, in the formula 2, A1 to A12 independently represent an amino acid; X does not exist, or is a moiety of 1- to 9-amino acid length; Helix B is a moiety of 11- to 13-amino acid length capable of forming an α-helix structure; and L is a moiety of 2- to 7-amino acid length represented by the formula 3, wherein, in the formula 3, the amino acids are numbered “i” (-1), “ii” (-2), and so on from the C-terminus side, provided that Liii to Lvii may not exist), and combination of three amino acids A1, A4 and Lii, or combination of two amino acids A4, and Lii is a combination corresponding to a target RNA base or base sequence.

主权项

1. A method for designing a protein that can bind to an RNA molecule in an RNA base-selective or RNA base sequence-specific manner, wherein:
the protein is a protein containing one or more of PPR motifs (preferably 2 to 14 PPR motifs) each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1:
[F1](HelixA)-X-(HelixB)-L  (Formula 1)(wherein:
Helix A is a moiety of 12-amino acid length capable of forming an α-helix structure, and is represented by the formula 2:
[F2]A1-A2-A3-A4-A5-A6-A7-A8-A9-A10-A11-A12  (Formula 2)wherein, in the formula 2, A1 to A12 independently represent an amino acid;
X does not exist, or is a moiety of 1- to 9-amino acid length; Helix B is a moiety of 11- to 13-amino acid length capable of forming an α-helix structure; and L is a moiety of 2- to 7-amino acid length represented by the formula 3;
[F3]Lvii-Lvi-Lv-Liv-Liii-Lii-Li  (Formula 3)wherein, in the formula 3, the amino acids are numbered “i” (-1), “ii” (-2), and so on from the C-terminus side,
provided that Liii to Lvii may not exist), and combination of three amino acids A1, A4 and Lii, or combination of two amino acids A4, and Lii is a combination corresponding to a target RNA base or base sequence.