Abstract
CRISPR-Cas9 has been adapted as readily programmable RNA-guided agents for genome engineering and manipulation. A major obstacle hampering its applications is the “off-target effect”, in which deleterious consequences arise from aberrant Cas9 activities on targets containing mismatch(es) within the 20-base-pair hybrid formed between the DNA protospacer and the RNA guide. RNA guide truncation has been developed as a viable strategy to reduce Cas9 off-target effects, although previous reports indicate that cell-based gene editing require a minimal guide length of 17 nucleotides (-nt) or longer.
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