CRISPR/CAS-related methods and compositions for treating cystic fibrosis

A CRISPR/Cas system, comprising:

• a) a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a cystic fibrosis transmembrane conductance regulator (CFTR) gene or a sodium channel epithelial 1 alpha (SCNN1A) gene, wherein said target sequence of said CFTR gene is within intron 2 or intron 10 of said CFTR gene, wherein said targeting domain that is complementary to said SCNN1A gene comprises a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a targeting domain sequence selected from the targeting domain sequences set forth in SEQ ID NOs:
• 497-654, 803-916, and 4176-15100 and wherein said targeting domain that is complementary to said CFTR gene comprises a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a targeting domain sequence selected from the targeting domain sequences set forth in SEQ ID NOs:
• 15101-27265;
• b) a Cas9 molecule;
• and
• c) a cDNA donor template that alters said CFTR gene or SCNN1A gene, wherein said cDNA donor template that alters said CFTR gene comprises (a) contiguous coding sequence of exon 3 through exon 27 of said CFTR gene;
• or (b) contiguous coding sequence of exon 11 through exon 27 of said CFTR gene.

A composition comprising:

• a) a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a CFTR gene or a SCNN1A gene, wherein the target sequence of the CFTR gene is within intron 2 or intron 10 of the CFTR gene, wherein said targeting domain that is complementary to said SCNN1A gene comprises a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a targeting domain sequence selected from the targeting domain sequences set forth in SEQ ID NOs:
• 497-654, 803-916, and 4176-15100 and wherein said targeting domain that is complementary to said CFTR gene comprises a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a targeting domain sequence selected from the targeting domain sequences set forth in SEQ ID NOs:
• 15101-27265;
• and
• b) a cDNA donor template that alters said CFTR gene or SCNN1A gene, wherein said cDNA donor template that alters said CFTR gene comprises (a) contiguous coding sequence of exon 3 through exon 27 of said CFTR gene;
• or (b) contiguous coding sequence of exon 11 through exon 27 of said CFTR gene.

A nucleic acid composition that comprises:

• a) a first nucleotide sequence that encodes a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a CFTR gene or a SCNN1A gene, wherein the target sequence of the CFTR gene is within intron 2 or intron 10 of the CFTR gene, wherein said targeting domain that is complementary to said SCNN1A gene comprises a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a targeting domain sequence selected from the targeting domain sequences set forth in SEQ ID NOs:
• 497-654, 803-916, and 4176-15100 and wherein said targeting domain that is complementary to said CFTR gene comprises a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a targeting domain sequence selected from the targeting domain sequences set forth in SEQ ID NOs:
• 15101-27265;
• and
• b) a template nucleic acid comprising a cDNA donor template that alters said CFTR gene or SCNN1A gene, wherein said cDNA donor template that alters said CFTR gene comprises (a) contiguous coding sequence of exon 3 through exon 27 of said CFTR gene;
• or (b) contiguous coding sequence of exon 11 through exon 27 of said CFTR gene.