Biallelic Knockout of PDCD1

A method for inactivating alleles of the Programmed cell death protein 1 (PDCD1) gene in a cell, the method comprising

• introducing to the cell a composition comprising:
• at least one CRISPR nuclease, or a polynucleotide molecule encoding a CRISPR nuclease;
• and
• an RNA molecule comprising a guide sequence portion, or a polynucleotide molecule encoding the RNA molecule,
• wherein a complex of the CRISPR nuclease and the RNA molecule affects a double strand break in alleles of the PDCD1 gene, and
• wherein the guide sequence portion of the RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs:
• 1-6220 modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully-complementary target sequence of the guide sequence portion.

A composition comprising:

• at least one CRISPR nuclease, or a polynucleotide molecule encoding a CRISPR nuclease;
• and
• an RNA molecule comprising a guide sequence portion, or a polynucleotide molecule encoding the RNA molecule,
• wherein the guide sequence portion of the RNA molecule comprises 17-50 contiguous nucleotides containing nucleotides in the sequence set forth in any one of SEQ ID NOs:
• 1-6220 or in any one of SEQ ID NOs:
• 1-6220, modified to contain 1, 2, 3, 4, or 5 nucleotide mismatches relative to a fully target sequence of the guide sequence portion.