MODULATING EXPRESSION OF POLYPEPTIDES VIA NEW GENE SWITCH EXPRESSION SYSTEMS
Disclosed herein are polynucleotides encoding ligand-inducible gene switch polypeptides, and systems comprising gene switch polypeptides for modulating the expression of a heterologous gene and an interleukin in a host cell. The compositions, methods and systems described herein facilitate ligand dependent expression of polypeptides including but not limited to cytokines and antigen binding polypeptides.
1 - 160 . (canceled)
161 . A polynucleotide comprising: (a) an AttP or AttB recombination sequence; and (b) a heterologous gene encoding a chimeric antigen receptor.
162 . The polynucleotide of claim 161 , comprising an AttP recombination sequence.
163 . The polynucleotide of claim 161 , comprising an AttB recombination sequence.
164 . The polynucleotide of claim 161 , wherein the chimeric antigen receptor is capable of binding CD19, CD33, BCMA, CD44, α-Folate receptor, CAIX, CD30, ROR1, CEA, EGP-2, EGP-40, HER2, HER3, Folate-binding Protein, GD2, GD3, IL-13R-a2, KDR, EDB-F, mesothelin, CD22, EGFR, MUC-1, MUC-16, MAGE-A1, h5T4, PSMA, TAG-72, EGFRvIII, CD123, and/or VEGF-R2.
165 . The polynucleotide of claim 161 , wherein the chimeric antigen receptor is capable of binding CD19, CD22, BCMA, EGFRvIII, CD33, ROR1, MUC-16, and/or mesothelin.
166 . The polynucleotide of claim 161 , wherein the chimeric antigen receptor comprises:
(a) an amino acid sequence having at least 90% identity with any one of SEQ ID NOs: 210, 211, 216, and 233-244;
(b) an amino acid sequence having at least 90% identity with SEQ ID NO: 214 and an amino acid sequence having at least 90% identity with SEQ ID NO: 215;
(c) an amino acid sequence having at least 90% identity with SEQ ID NO: 220 and an amino acid sequence having at least 90% identity with SEQ ID NO: 221;
(d) an amino acid sequence having at least 90% identity with SEQ ID NO: 222 and an amino acid sequence having at least 90% identity with SEQ ID NO: 223;
(e) an amino acid sequence having at least 90% identity with SEQ ID NO: 224 and an amino acid sequence having at least 90% identity with SEQ ID NO: 225; or
(f) an amino acid sequence having at least 90% identity with SEQ ID NO: 226 and an amino acid sequence having at least 90% identity with SEQ ID NO: 227.
167 . The polynucleotide of claim 161 , further encoding a cell tag.
168 . The polynucleotide of claim 167 , wherein the cell tag comprises an amino acid sequence having at least 90% identity with any one of SEQ ID NOs: 190-195 and 197-200.
169 . The polynucleotide of claim 167 , wherein the cell tag comprises a sequence having at least 90% identity with SEQ ID NO: 190.
170 . The polynucleotide of claim 161 , further encoding a cytokine.
171 . The polynucleotide of claim 170 , wherein the cytokine is IL-1, IL-2, IL-12, IL-15, or IL-21, or a functional variant thereof.
172 . The polynucleotide of claim 170 , wherein the cytokine is a membrane-bound cytokine.
173 . The polynucleotide of claim 172 , wherein the membrane-bound cytokine is membrane-bound IL-15 (mbIL-15).
174 . The polynucleotide of claim 161 , further encoding a fusion protein comprising: (a) IL-15, or a functional variant thereof; and (b) IL-15Rα, or a functional variant thereof.
175 . The polynucleotide of claim 174 , wherein the fusion protein comprises a sequence having at least 90% identity with SEQ ID NO: 205.
176 . The polynucleotide of claim 161 , further encoding a serine recombinase.
177 . The polynucleotide of claim 176 , wherein the serine recombinase is a SF370, SPβc2, Bxb1, A118, or φRv1 serine recombinase.
178 . The polynucleotide of claim 176 , wherein the serine recombinase is an SF370 recombinase.
179 . The polynucleotide of claim 176 , wherein the serine recombinase is an SPβc2 recombinase.
180 . A vector comprising the polynucleotide of claim 161 .