Randomized configuration targeted integration of nucleic acids
AssigneeGenentech, Inc.
InventorsShahram Misaghi, Bradley R. Snedecor
Life of this patent
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Abstract
The presently disclosed subject matter relates to “Randomized Configuration Targeted Integration” (also referred to herein as “Randomized Chain Targeted Integration”) (RCTI) strategies for the generation and identification of host cells capable of expressing recombinant proteins, e.g., monoclonal antibodies, as well as compositions derived from the same, e.g., bispecific antibodies, and other complex format proteins, e.g., membrane protein complexes and other difficult to express molecules.
Claims49
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 006874047.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 006884592.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 006881296.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 003616412.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 003615063.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 006882936.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
A method for generating and high throughput screening a library of targeted integration (TI) host cells expressing at least one sequence of interest (SOI) comprising:
- a) generating a library of TI host cells, wherein said library comprises a plurality of TI host cells expressing one or more SOIs, by:
- i) providing a plurality of TI host cells;
- ii) contacting the plurality of TI host cells with a plurality of vectors, wherein the vectors comprise one or more SOIs;
- iii) introducing the one or more SOIs into a locus of the genome of the one or more of the plurality of TI host cells wherein the locus is at least about 90% homologous to the sequence of NW 003615411.1;
- b) separating said library into single clones;
- and
- c) screening the clones for a specific cellular or product attribute.
Dependencies are inferred from the claim text.