Compositions and methods for modifying a target nucleic acid
AssigneesThe Regents of the University of California, The J. David Gladstone Institutes, a testamentary trust established under the Will of J. David Gladstone
InventorsAlexander Marson, Brian Shy, Justin Eyquem, Vivasvan Vykunta
Life of this patent
Estimating the term…
Abstract
The disclosure provides compositions and methods for for modifying an endogenous cell surface protein (e.g., an endogenous TCR) in a cell (e.g., a T cell) with a CAR or an exogenous protein (e.g., an exogenous intracellular or cell surface protein (e.g., an exogenous TCR)).
Claims7
A method for modifying an endogenous T cell receptor alpha constant chain (TRAC) genomic locus in a T cell with at least two chimeric antigen receptors (CARs), the method comprising the step of introducing into the T cell:
- i) a guide RNA (gRNA) comprising the sequence of CTGGATATCTGTGGGACAAG (SEQ ID NO:3), ATCTGTGGGACAAGAGGATC (SEQ ID NO:4), TCTGTGGGACAAGAGGATCA (SEQ ID NO:5), GGGACAAGAGGATCAGGGTT (SEQ ID NO:6), TCTTTGCCCCAACCCAGGCT (SEQ ID NO:7), CTTTGCCCCAACCCAGGCTG (SEQ ID NO:8), TGGAGTCCAGATGCCAGTGA (SEQ ID NO:9), ACTACCGTTTACTCGATATA (SEQ ID NO:
- 17), TCGAGTAAACGGTAGTGCTG (SEQ ID NO:
- 18), TAGTGCTGGGGCTTAGACGC (SEQ ID NO:
- 19), ATGGGAGGTTTATGGTATGT (SEQ ID NO:20), CTGGGCATTAGCAGAATGGG (SEQ ID NO:21), CTAATGCCCAGCCTAAGTTG (SEQ ID NO:22), GTACATCTTGGAATCTGGAG (SEQ ID NO:23), AACTCTGGCAGAGTAAAGGC (SEQ ID NO:24), CTGCCAGAGTTATATTGCTG (SEQ ID NO:25), GTGAACGTTCACTGAAATCA (SEQ ID NO:26), AGCTATCAATCTTGGCCAAG (SEQ ID NO:27), or CAGGCACAAGCTATCAATCT (SEQ ID NO:28);
- ii) a homology-directed repair template (HDRT) comprising a sequence having at least 95% identity to the sequence of SEQ ID NO:10 or 11 encoding the at least two CARs;
- and
- iii) a Cas protein,
- thereby integrating the nucleotide sequence encoding the at least two CARs into the endogenous TRAC locus of the T cell.
A composition comprising:
- i) a guide RNA (gRNA) comprising the sequence of CTGGATATCTGTGGGACAAG (SEQ ID NO:3), ATCTGTGGGACAAGAGGATC (SEQ ID NO:4), TCTGTGGGACAAGAGGATCA (SEQ ID NO:5), GGGACAAGAGGATCAGGGTT (SEQ ID NO:6), TCTTTGCCCCAACCCAGGCT (SEQ ID NO:7), CTTTGCCCCAACCCAGGCTG (SEQ ID NO:8), TGGAGTCCAGATGCCAGTGA (SEQ ID NO:9), ACTACCGTTTACTCGATATA (SEQ ID NO:
- 17), TCGAGTAAACGGTAGTGCTG (SEQ ID NO:
- 18), TAGTGCTGGGGCTTAGACGC (SEQ ID NO:
- 19), ATGGGAGGTITTATGGTATGT (SEQ ID NO:20), CTGGGCATTAGCAGAATGGG (SEQ ID NO:21), CTAATGCCCAGCCTAAGTTG (SEQ ID NO:22), GTACATCTTGGAATCTGGAG (SEQ ID NO:23), AACTCTGGCAGAGTAAAGGC (SEQ ID NO:24), CTGCCAGAGTTATATTGCTG (SEQ ID NO:25), GTGAACGTTCACTGAAATCA (SEQ ID NO:26), AGCTATCAATCTTGGCCAAG (SEQ ID NO:27), or CAGGCACAAGCTATCAATCT (SEQ ID NO:28);
- and
- ii) a homology-directed repair template (HDRT) comprising a sequence having at least 95% identity to the sequence of SEQ ID NO:10 or 11.
Dependencies are inferred from the claim text.