Systems and methods for one-shot guide RNA (ogRNA) targeting of endogenous and source DNA

An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a Cpf1 protein, wherein the nucleic acid sequence comprises a eukaryotic nucleic acid sequence that

• a) is at least 17 nucleotides in length;
• and
• b) comprises or is adjacent to a protospacer adjacent motif (PAM) that is recognized by the Cpf1 protein,
• wherein the Cpf1 comprises an amino acid insertion, relative to SEQ ID NO:
• 13, at a position selected from the group consisting of:
• i) between amino acid positions 147 and 148;
• ii) anywhere between amino acid positions 484 and 492,
• iii) anywhere between amino acid positions 568 and 590,
• iv) anywhere between amino acid positions 795 and 855,
• v) anywhere between amino acid positions 1131 and 1140, and
• vi) anywhere between amino acid positions 1160 and 1173, and
• where the insertion relative to SEQ ID NO:
• 13 results in a Cpf1 protein having at least 95% sequence identity to SEQ ID NO:
• 13.