The University of Western Australia

Medical Research Council

Oliver Rackham

Aleksandra Filipovska

Jason W Chin

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 ssDNA-binding domain which itself comprises at least a pair of PPR ssDNA base-binding motifs. The PPR ssDNA base-binding motifs of the PPR ssDNA-binding domain are operably capable of binding the target ssDNA. Recombinant polypeptides comprising at least one PPR sDNA-binding domain capable of binding to target ssDNA sequence are also described, together with fusion proteins comprising the recombinant PPR ssDNA-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.

Proteins and their use for nucleotide binding

A recombinant polypeptide is described which comprises at least one PUF RNA-binding domain capable of specifically binding to a cytosine RNA base. The PUF RNA-binding domain of the polypeptide comprises at least one RNA base-binding motif of the general formula XXXXXXXXXXXwherein Xis selected from the group including glutamine (Q), valine (V), methionine (M), proline (P), glutamic acid (E), and lysine (K); Xis selected from the group including histidine (H), phenylalanine (F), tyrosine (Y), and asparagine (N); Xis selected from the group including glycine (G) and alanine (A); Xis selected from the group including glycine (G), alanine (A), serine (S), threonine (T) and cysteine (C); Xis selected from the group including arginine (R), tyrosine (Y), histidine (H), and asparagine (N); Xis selected from the group including phenylalanine (F), leucine (L), and valine (V); Xis selected from the group including isoleucine (I), leucine (L), and valine (V); Xis arginine (R); Xis selected from the group including leucine (L), lysine (K), arginine (R), glutamine (Q), and histidine (H); Xis selected from the group including lysine (K), phenylalanine (F), alanine (A), cysteine (C), isoleucine (I), valine (V), leucine (L), and methionine (M); and Xis selected from the group including leucine (L), phenylalanine (F), isoleucine (I), and valine (V); and wherein the RNA base-binding motif is operably capable of specifically binding to a cytosine RNA base.

Peptides for the specific binding of RNA targets

A recombinant polypeptide is described including at least one PUF RNA-binding domain capable of specifically binding to a cytosine RNA base. The PUF RNA-binding domain of the polypeptide includes at least one RNA base-binding motif of the general formula XXXXXXXXXXXwherein Xis selected from a defined group and wherein the RNA base-binding motif is operably capable of specifically binding to a cytosine RNA base.

Orthogonal ribosome orthogonal mRNA pairs are provided, as are methods for their selection involving a novel positive-negative selection approach, and methods for their use. Also provided are cellular logic circuits involving orthogonal ribosomes.

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Western Australia, Australia

Oliver Rackham