US 2026/0209747 A1 · DIRECTED EVOLUTION OF ENGINEERED VIRUS-LIKE PARTICLES (EVLPS)

Abstract

The present disclosure provides methods, compositions, and systems for evolving virus-like particles (VLPs) having one or more desired properties such as increased production levels, increased cargo packaging efficiency, and/or increased transduction of particular target cell types of interest. The present disclosure also provides libraries for use in such methods, and methods for producing the libraries. Group specific antigen (gag) proteins comprising nucleocapsid protein variants evolved using the methods described herein are also provided herein. The present disclosure also provides VLPs comprising such gag proteins comprising nucleocapsid protein variants. Polynucleotides, vectors, cells, and kits useful for performing the methods described herein are also provided.

Claims32

Dependent claims

A method for generating a library of cells capable of producing virus-like particles (VLPs) comprising:

transfecting a plurality of polynucleotides encoding VLPs into producer cells,
wherein each polynucleotide encodes a variant of at least one component of the VLP or its cargo,
wherein the variant comprises at least one amino acid substitution relative to a VLP that does not comprise a variant of at least one component of the VLP or its cargo;
and
wherein each polynucleotide encoding a VLP component or cargo variant is associated with a unique nucleic acid barcode sequence.

2 more elements

A group specific antigen (gag) protein comprising a viral nucleocapsid protein variant that comprises an amino acid sequence at least 80% identical to the amino acid sequence of SEQ ID NO:

100, wherein the amino acid sequence of the viral nucleocapsid protein variant comprises one or more substitutions at positions selected from the group consisting of 215, 216, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 244, 245, 246, 249, 250, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 269, 271, 272, 274, 275, 276, 277, 279, 280, 281, 282, 283, 285, 286, 288, 289, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 307, 308, 310, 311, 418, 420, 421, 424, 427, 430, 432, 433, 435, 436, 437, 438, 440, 441, 443, 444, 446, 447, 448, 449, 452, 455, 458, 460, 463, 464, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 477, 478, 479, 480, 481, 482, 483, 485, 486, 487, 488, 489, 490, 491, 492, 495, 496, 497, 498, 499, 500, 501, 502, 503, 504, 505, 506, 507, 508, 509, 510, 511, and 512 relative to the amino acid sequence provided in SEQ ID NO:
100, or at corresponding positions in a homologous viral nucleocapsid protein.

Dependencies are inferred from the claim text.

Directed evolution of engineered virus-like particles (evlps)

Life of this patent

Directed evolution of engineered virus-like particles (evlps)

Life of this patent

Abstract

Claims32