GUIDE STRAND LIBRARY CONSTRUCTION AND METHODS OF USE THEREOF

摘要

An improved method for the rapid and efficient production of guide strand libraries is disclosed. Also included are kits comprising reagents suitable for practicing the method. Embodiments may include a polynucleotide comprising a sequence encoding for a constant region comprising a protein binding segment of an RNA component of a CRISPR complex. The sequence may comprise a non-palindromic recognition sequence for a first restriction enzyme having methyltransferase activity and configured to cleave at least 17 nucleotides outside of the non-palindromic recognition sequence, wherein at least one base of the non-palindromic recognition sequence is configured to be methylated by the first restriction enzyme, thereby blocking endonuclease activity of the first restriction enzyme.