	US 11,742,056 B2
	Method for designing RNA-binding protein utilizing PPR motif, and use thereof
Takahiro Nakamura, Fukuoka (JP); Yusuke Yagi, Fukuoka (JP); and Keiko Kobayashi, Fukuoka (JP)
Assigned to KYUSHU UNIVERSITY, NATIONAL UNIVERSITY CORPORATION, Fukuoka (JP)
Filed by KYUSHU UNIVERSITY, NATIONAL UNIVERSITY CORPORATION, Fukuoka (JP)
Filed on Mar. 8, 2021, as Appl. No. 17/195,449.
Application 17/195,449 is a division of application No. 16/894,295, filed on Jun. 5, 2020, granted, now 10,943,671, issued on Mar. 9, 2021.
Application 16/894,295 is a division of application No. 16/431,429, filed on Jun. 4, 2019, granted, now 10,679,731, issued on Jun. 9, 2020.
Application 16/431,429 is a division of application No. 15/962,127, filed on Apr. 25, 2018, granted, now 10,340,028, issued on Jul. 2, 2019.
Application 15/962,127 is a division of application No. 15/335,243, filed on Oct. 26, 2016, granted, now 9,984,202, issued on May 29, 2018.
Application 15/335,243 is a division of application No. 14/352,697, granted, now 9,513,283, issued on Dec. 6, 2016, previously published as PCT/JP2012/077274, filed on Oct. 22, 2012.
Claims priority of application No. 2011-231346 (JP), filed on Oct. 21, 2011.
Prior Publication US 2021/0249103 A1, Aug. 12, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. G16B 15/00 (2019.01); G16B 30/00 (2019.01); G16B 35/00 (2019.01); G16C 20/60 (2019.01); C07K 14/415 (2006.01); C12N 15/82 (2006.01); G16B 20/50 (2019.01); G16B 20/30 (2019.01); G16B 30/10 (2019.01); C12Q 1/68 (2018.01); G01N 33/53 (2006.01); G16B 20/00 (2019.01)

CPC G16B 15/00 (2019.02) [C07K 14/415 (2013.01); C12N 15/8216 (2013.01); C12N 15/8287 (2013.01); C12N 15/8289 (2013.01); C12Q 1/68 (2013.01); G01N 33/5308 (2013.01); G16B 20/30 (2019.02); G16B 20/50 (2019.02); G16B 30/00 (2019.02); G16B 30/10 (2019.02); G16B 35/00 (2019.02); G16C 20/60 (2019.02); C07K 2319/85 (2013.01); C12Q 2522/101 (2013.01); G16B 20/00 (2019.02)]

20 Claims

1. A method for controlling a function of an RNA having a target sequence, comprising:

determining a nucleic acid sequence coding for an amino acid sequence of a protein,

cloning said nucleic acid sequence, and

preparing a vector carrying a polynucleotide consisting of the nucleic acid sequence,

wherein the protein contains one or more of PPR motifs, each consisting of a polypeptide of 30- to 38-amino acid length represented by the formula 1:

(HelixA)-X-(HelixB)-L (Formula 1)

wherein, in formula 1, Helix A is a moiety of 12-amino acid length capable of forming an α-helix structure, and is represented by the formula 2:

A₁-A₂-A₃-A₄-A₅-A₆-A₇-A₈-A₉-A₁₀-A₁₁-A₁₂ (Formula 2)

wherein, in the formula 2, A₁to A₁₂independently represent an amino acid;

wherein, in the formula 1, X is a moiety of 1- to 9-amino acid length and is optional;

wherein, in the formula 1, Helix B is a moiety of 11- to 13-amino acid length capable of forming an α-helix structure; and

wherein, in the formula 1, L is a moiety of 2- to 7-amino acid length represented by the formula 3;

L_vii-L_vi-L_v-L_iv-L_iii-L_ii-L_i (Formula 3)

wherein, in the formula 3, L_ito L_viindependently represent an amino acid, and L_iiito L_viare optional, and

wherein a combination of three amino acids A₁, A₄and L_ii, or a combination of two amino acids A₄and L_iicorresponds to the target sequence;

preparing a cell containing the RNA having the target sequence; and

introducing the vector into the cell, whereby the protein binds to the RNA having the target sequence.