US 2022/0025381 A1 · INDUCIBLE CONTROL OF GENE EXPRESSION

Abstract

The technology described herein is directed to inducible and repressible polypeptides and polypeptide systems. In particular, described herein are split sequence-specific nucleases and split recombinases that are linked to drug-inducible or drug-repressible dimerization domains. In some embodiments, the polypeptides comprise sequestering domains and/or have their expression controlled by an inducible promoter. In multiple aspects described herein are polynucleotides, vectors, cells, and pharmaceutical compositions comprising said polypeptides or polypeptide systems. Also described herein are methods of using said polypeptide systems to modulate the expression of a target polypeptide or to treat a subject in need of a cell therapy.

Claims30

Dependent claims

An inducible split-nuclease polypeptide system comprising:

a) a first polypeptide comprising:
i) a first member of an inducible dimerization domain (D1);
and
ii) a first polypeptide fragment of a sequence-specific nuclease (N1);
and
b) a second polypeptide comprising:
i) a second member of the inducible dimerization domain (D2);
and
ii) a second polypeptide fragment of the sequence-specific nuclease (N2);
and
wherein the first and second members of the inducible dimerization domain come together in the presence of an inducer agent or inducer signal, resulting in protein complementation of the two nuclease polypeptide fragments to form the active nuclease protein in the presence of the inducer agent or inducer signal.

8 more elements

An inducible split-recombinase polypeptide system comprising:

a) a first polypeptide comprising:
i) a first member of an inducible dimerization domain (D1);
and
ii) a first polypeptide fragment of a recombinase (R1);
and
b) a second polypeptide comprising:
i) a second member of the inducible dimerization domain (D2);
and
ii) a second polypeptide fragment of the recombinase (R2);
and
wherein the first and second members of the inducible dimerization domain come together in the presence of an inducer agent or inducer signal, resulting in protein complementation of the two recombinase polypeptide fragments to form the active recombinase protein in the presence of the inducer agent or inducer signal.

8 more elements

A repressible split-nuclease polypeptide system comprising:

a) a first polypeptide comprising:
i) a first member of a repressible dimerization domain (RD1);
and
ii) a first polypeptide fragment of a sequence-specific nuclease (N1);
and
b) a second polypeptide comprising:
i) a second member of the repressible dimerization domain (RD2);
and
ii) a second polypeptide fragment of the sequence-specific nuclease (N2);
and
wherein the first and second members of the repressible dimerization domain come together in the absence of a repressor agent or target signal, resulting in protein complementation of the two nuclease polypeptide fragments to form the active nuclease protein in the absence of the repressor agent or repressor signal;
wherein the first and second members of the repressible dimerization domain do not come together in the presence of the repressor agent or repressor signal, resulting in no protein complementation of the two nuclease polypeptide fragments and no formation of the active nuclease protein in the presence of the repressor agent or repressor signal.

9 more elements

A repressible split-recombinase polypeptide system comprising:

a) a first polypeptide comprising:
i) a first member of a repressible dimerization domain (RD1);
and
ii) a first polypeptide fragment of a recombinase (R1);
and
b) a second polypeptide comprising:
i) a second member of the repressible dimerization domain (RD2);
and
ii) a second polypeptide fragment of the recombinase (R2);
and
wherein the first and second members of the repressible dimerization domain come together in the absence of a repressor agent or repressor signal, resulting in protein complementation of the two recombinase polypeptide fragments to form the active recombinase protein in the absence of the repressor agent or repressor signal;
wherein the first and second members of the repressible dimerization domain do not come together in the presence of the repressor agent or repressor signal, resulting in no protein complementation of the two recombinase polypeptide fragments and no formation of the active recombinase protein in the presence of the repressor agent or repressor signal.

9 more elements

Dependencies are inferred from the claim text.

Inducible control of gene expression

Life of this patent

Inducible control of gene expression

Life of this patent

Abstract

Claims30