Self-inactivating endonuclease-encoding nucleic acids and methods of using the same

A recombinant expression vector comprising:

• i) a nucleotide sequence encoding a CRISPR-Cas endonuclease;
• ii) a regulatory element operably linked to the CRISPR-Cas endonuclease-encoding nucleotide sequence;
• and
• iii) flanking nucleotide sequences flanking the CRISPR-Cas endonuclease-encoding nucleotide sequence, wherein the flanking nucleotide sequences flanking the CRISPR-Cas endonuclease-encoding nucleotide sequence are cleavable by the CRISPR-Cas endonuclease when the CRISPR-Cas endonuclease is complexed with a guide RNA comprising:
• a) a CRISPR-Cas endonuclease binding segment that binds to the CRISPR-Cas endonuclease and b) a guide sequence that hybridizes to:
• (i) a target host nucleotide sequence in a target nucleic acid of a host cell;
• and (ii) the flanking nucleotide sequences,
• wherein each of the flanking nucleotide sequences has a length of 15 or more nucleotides and comprises:
• (i) 5 or fewer mismatches with the target host nucleotide sequence;
• or (ii) at least 70% sequence identity to the target host nucleotide sequence,
• wherein the recombinant expression vector is configured to express the CRISPR-Cas endonuclease in the host cell,
• and wherein cleavage of the recombinant expression vector by the CRISPR-Cas endonuclease complexed with the guide RNA reduces expression of the CRISPR-Cas endonuclease in the host cell.