The genetic information encoded in the reverse-transcription template (RTT) can then be copied into the targeted site. Upon binding, the Cas9 nickase nicks the target strand and the reverse transcriptase uses the primer binding site (PBS) of the pegRNA to initiate the reverse transcription. S1) are a Cas9 nickase fused with reverse transcriptase and a prime editing guide RNA (pegRNA), which brings the PE machinery to targeted sites through a standard single-guide RNA (sgRNA) sequence. The key components of a prime editor (PE) (Additional file 1: Fig. Prime editing, a newly invented genome-editing technology, enables all types of base transitions and transversions to be accomplished, as well as customized insertions (up to 44 nucleotides) and deletions (up to 80 nucleotides). However, base editors are not suitable for generating other types of point mutations or for insertions and deletions. In contrast, base editors can generate transition point mutations with high efficiency and accuracy without introducing double-strand breaks. Standard CRISPR-Cas9 approaches tend to introduce imprecise edits with indels varying in size from a single nucleotide to hundreds of nucleotides through nonhomologous end joining (NHEJ). Different CRISPR-based systems have their own strengths and weaknesses. Among different genome-editing technologies, the clustered regularly interspaced short palindromic repeats (CRISPR)–based systems are the most widely used ones. Genome-editing technologies have revolutionized genetic studies ranging from those involving traditional interventions to precise manipulations of DNA sequences, offering both simplicity and robust outcomes.