Rational design of minimum CRISPR guide RNA by site-specific Cas9-RNA conjugation.

The CRISPR–Cas9 system enables facile and efficient genome engineering in living cells and organisms. We report a Cas9–RNA conjugation strategy to afford minimal crRNA containing only the guide sequence for the target gene, which may simplify and reduce the cost for large-scale and high-throughput crRNA synthesis and lead to new insights into the design of CRISPR family complexes for diverse purposes.