US 2022/0220166 A1Published application

Ppr protein causing less aggregation and use of the same

AssigneesEditForce, Inc., Kyushu University, National University Corporation

InventorsYusuke Yagi, Takayoshi Imai, Takayuki Tamai, Takahiro Nakamura, Takamasa Teramoto

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Abstract

In order to improve aggregation property of a PPR protein, the A6 amino acid of the 1st PPR motif from the N-terminus (M1) is made more hydrophilic. Further, the A9 amino acid of M1 is made to be a hydrophilic amino acid or glycine. The A6 amino acid is preferably asparagine or aspartic acid, and the A9 amino acid is preferably glutamine, glutamic acid, lysine, or glycine. Proteins containing such a PPR motif as M1 motif may have not only improved aggregation property, but also high binding power to a target nucleic acid.

Claims18

Dependent claims

A PPR motif, which is any one of the following PPR motifs:

(C-1) a PPR motif consisting of any one of the sequences of SEQ ID NOS:
4 to 7;
(C-2) a PPR motif consisting of any one of the sequences of SEQ ID NOS:
4 to 7 having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having a cytosine-binding property;
(C-3) a PPR motif having a sequence identity of at least 80% to any one of the sequences of SEQ ID NOS:
4 to 7, provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having a cytosine-binding property;
(A-1) a PPR motif consisting of the sequence of SEQ ID NO:
8 having a substitution of the amino acid at position 6 with asparagine or aspartic acid:
(A-2) a PPR motif consisting of the sequence of (A-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having an adenine-binding property;
(A-3) a PPR motif having a sequence identity of at least 80% to the sequence of (A-1), provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having an adenine-binding property;
(G-1) a PPR motif consisting of the sequence of SEQ ID NO:
9 having a substitution of the amino acid at position 6 with asparagine or aspartic acid;
(G-2) a PPR motif consisting of the sequence of (G-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having a guanine-binding property;
(G-3) a PPR motif having a sequence identity of at least 80% to the sequence of (G-1), provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having a guanine-binding property;
(U-1) a PPR motif consisting of the sequence of SEQ ID NO:
10 having a substitution of the amino acid at position 6 with asparagine or aspartic acid;
(U-2) a, PPR motif consisting of the sequence of (U-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having a uracil-binding property;
and
(U-3) a PPR motif having a sequence identity of at least 80% to the sequence of (U-1), provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having a uracil-binding property.

16 more elements

A PPR motif, which is any one of the following PPR motifs:

(C-1) a PPR motif consisting of any one of the sequences of SEQ ID NOS:
4 to 7;
(C-2) a PPR motif consisting of any one of the sequences of SEQ ID NOS:
4 to 7 having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, 9, and 34, and having a cytosine-binding property;
(C-3) a PPR motif having a sequence identity of at least 80% to any one of the sequences of SEQ ID NOS:
4 to 7, provided that the amino acids at positions 1, 4, 6, 9, and 34 are identical, and having a cytosine-binding property;
(A-1) a PPR motif consisting of the sequence of SEQ ID NO:
8 having such substitutions of the amino acids at positions 6 and 9 that any one of the combinations defied below is satisfied;
(A-2) a PPR motif consisting of the sequence of (A-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, 9, and 34, and having an adenine-binding property;
(A-3) a PPR motif having a sequence identity of at least 80% to the sequence of (A-1), provided that the amino acids at positions 1, 4, 6, 9, and 34 are identical, and having an adenine-binding property;
(G-1) a PPR motif consisting of the sequence of SEQ ID NO:
9 having such substitutions of the amino acids at positions 6 and 9 that any one of the combinations defined below is satisfied;
(G-2) a PPR motif consisting of the sequence of (G-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, 9, and 34, and having a guanine-binding property;
(G-3) a PPR motif having a sequence identity of at least 80% to the sequence of((G-1), provided that the amino acids at positions 1, 4, 6, 9, and 34 are identical, and having a guanine-binding property;
(U-1) a PPR motif consisting of the sequence of SEQ ID NO:
10 having such substitutions of the amino acids at positions 6 and 9 that any one of the combinations defined below is satisfied:
(U-2) a PPR motif consisting of the sequence of (U-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, 9, and 34, and having a uracil-binding property;
and
(U-3) a PPR motif having a sequence identity of at least 80% to the sequence of (U-1), provided that the amino acids at positions 1, 4, 6, 9, and 34 are identical, and having a uracil-binding property.
a combination of asparagine as the amino acid at position 6 and glutamic acid as the amino acid at position 9,
a combination of asparagine as the amino acid at position 6 and glutamine as the amino acid at position 9,
a combination of asparagine as the amino acid at position 6 and lysine as the amino acid at position 9, and
a combination of aspartic acid as the amino acid at position 6 and glycine as the amino acid at position 9.

20 more elements

Dependencies are inferred from the claim text.

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Abstract

A PPR motif, which is any one of the following PPR motifs:

(C-1) a PPR motif consisting of any one of the sequences of SEQ ID NOS:
4 to 7;
(C-2) a PPR motif consisting of any one of the sequences of SEQ ID NOS:
4 to 7 having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having a cytosine-binding property;
(C-3) a PPR motif having a sequence identity of at least 80% to any one of the sequences of SEQ ID NOS:
4 to 7, provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having a cytosine-binding property;
(A-1) a PPR motif consisting of the sequence of SEQ ID NO:
8 having a substitution of the amino acid at position 6 with asparagine or aspartic acid:
(A-2) a PPR motif consisting of the sequence of (A-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having an adenine-binding property;
(A-3) a PPR motif having a sequence identity of at least 80% to the sequence of (A-1), provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having an adenine-binding property;
(G-1) a PPR motif consisting of the sequence of SEQ ID NO:
9 having a substitution of the amino acid at position 6 with asparagine or aspartic acid;
(G-2) a PPR motif consisting of the sequence of (G-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having a guanine-binding property;
(G-3) a PPR motif having a sequence identity of at least 80% to the sequence of (G-1), provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having a guanine-binding property;
(U-1) a PPR motif consisting of the sequence of SEQ ID NO:
10 having a substitution of the amino acid at position 6 with asparagine or aspartic acid;
(U-2) a, PPR motif consisting of the sequence of (U-1) having a substitution, deletion, or addition of 1 to 9 amino acids other than the amino acids at positions 1, 4, 6, and 34, and having a uracil-binding property;
and
(U-3) a PPR motif having a sequence identity of at least 80% to the sequence of (U-1), provided that the amino acids at positions 1, 4, 6, and 34 are identical, and having a uracil-binding property.

16 more elements