PEPTIDES FOR THE BINDING OF NUCLEOTIDE TARGETS

摘要

A method of regulating expression of a gene in a cell is described, comprising the step of introducing into the cell a recombinant polypeptide comprising a PPR RNA-binding domain which itself comprises at least a pair of PPR RNA base-binding motifs. The PPR RNA base-binding motifs of the PPR RNA-binding domain are operably capable of binding the target RNA molecule with a target RNA sequence. Recombinant polypeptides comprising at least one PPR RNA-binding domain capable of binding to target RNA sequence are also described, together with fusion proteins comprising the recombinant PPR RNA-binding domains as well as isolated nucleic acids useful in preparing the recombinant polypeptides described. Recombinant vectors; compositions comprising the recombinant polypeptides; isolated nucleic acids; recombinant vectors; host cells comprising same; use of same in the manufacture of a medicament for regulating gene expression; as well as systems and kits for regulating gene expression are also described.

主权项

1. A recombinant polypeptide comprising at least one PPR RNA-binding domain capable of binding to a target RNA sequence, the PPR RNA-binding domain comprising at least two PPR RNA base-binding motifs comprising
a.
i. amino acid position six of a first PPR RNA base-binding motif comprises threonine (T), serine (S), or glycine (G);ii. amino acid position one of a second adjacent PPR binding motif comprises asparagine (N), threonine (T), or serine (S); andiii. the PPR domain is operably capable of binding to an adenine (A) RNA base in a target RNA sequence; b.
i. amino acid position six of a first PPR RNA base-binding motif comprises threonine (T), serine (S), glycine (G), or alanine (A);ii. amino acid position one of a second adjacent PPR binding motif comprises aspartic acid (D), threonine (T), or serine (S); andiii. the PPR domain is operably capable of binding to a guanine (G) RNA base in a target RNA sequence; c.
i. amino acid position six of a first PPR RNA base-binding motif comprises threonine (T) or asparagine (N);ii. amino acid position one of a second adjacent PPR binding motif comprises asparagine (N), serine (S), aspartic acid (D), or threonine (T); andiii. the PPR domain is operably capable of binding to a cytosine (C) RNA base in a target RNA sequence; and d.
i. amino acid position six of a first PPR RNA base-binding motif comprises threonine (T) or asparagine (N);ii. amino acid position one of a second adjacent PPR binding motif comprises aspartic acid (D), serine (S), asparagine (N), or threonine (T); andiii. the PPR domain is operably capable of binding to a uracil (U) RNA base in a target RNA sequence.