Customized class switch of immunoglobulin genes in lymphoma and hybridoma by CRISPR/CAS9 technology
AssigneeChildren's Medical Center Corporation
InventorsRoberto Chiarle, Taek-Chin Cheong
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Abstract
The present disclosure provides genetically modified antibody-producing cells comprising edited chromosomal sequences associated with immunoglobulin heavy chain constant region, the IgH locus. In particular, these cells are generated using a CRISPR/Cas 9-mediated editing process. The disclosure also provides specific guide RNA (gRNA) guide sequences that target the chromosomal sequence of immunoglobulin heavy chain constant region in the Switch regions.
Claims16
A composition for directing class switch recombination (CSR) of an immunoglobulin heavy (IgH) chain in a mammalian cell to a different IgH subclass, the composition comprising a vector comprising a nucleic acid encoding a Cas9 nuclease or nickase and nucleic acids encoding at least two guide RNAs (gRNA), wherein the at least two gRNA are selected from SEQ ID NOs 5-8, 11-15, 56-65, 68-223, and 257-399.
A method for directing class switch recombination (CSR) of the immunoglobulin heavy (IgH) chain in a mammalian cell to a different IgH subclass comprising contacting the mammalian cell with a vector comprising a nucleic acid encoding a Cas9 nuclease or nickase and nucleic acids encoding at least two guide RNAs (gRNA), or contacting with a composition comprising said vector, wherein the at least two gRNA is selected from SEQ ID NOs 5-8, 11-15, 56-65, 68-223, and 257-399.
Dependencies are inferred from the claim text.