Abstract: Disclosed are methods of targeted sequencing by using gRNA-endonuclease complexes and methods of designing pools of gRNAs. The disclosure also provides sequencing adapters that that comprise a double-stranded nucleic acid having a single-stranded overhang with degenerate overhanging bases. In a first aspect, methods for Dephosphorylate targeted sequencing of double-stranded nucleic acids comprises cleaving dephosphorylated double-stranded nucleic acids with a plurality of endonuclease-guide ribonucleic acid (gRNA) complexes to generate double-stranded nucleic acid fragments having phosphorylated 5? end overhangs at targeted sites.