摘要
The development of CRISPR-Cas9 based genetic manipulation tools represents a huge breakthrough in life sciences and has been stimulating research on metabolic engineering,synthetic biology,and systems biology.The CRISPR-Cas9 and its derivative tools are one of the best choices for precise genome editing,multiplexed genome editing,and reversible gene expression control in microorganisms.However,challenges remain for applying CRISPR-Cas9 in novel microorganisms,especially those industrial microorganism hosts that are intractable using traditional genetic manipulation tools.How to further extend CRISPR-Cas9 to these microorganisms is being an urgent matter.In this review,we first introduce the mechanism and application of CRISPR-Cas9,then discuss how to optimize CRISPR-Cas9 as genome editing tools,including but not limited to how to reduce off-target effects and Cas9 related toxicity,and how to increase on-target efficiency by optimizing crRNA and sgRNA design.
基金
This work was supported by the National Key R&D Program of China(2018YFA0901600)
the National Natural Science Foundation of China(31900052)
Youth Innovation Promotion Association,Chinese Academy of Sciences(2020182).
