초록
<P/><P><I><I>Corynebacterium glutamicum</I></I> is an important organism for the industrial production of amino acids. Metabolic pathways in this organism are usually engineered by conventional methods such as homologous recombination, which depends on rare double-crossover events. To facilitate the mapping of gene expression levels to metabolic outputs, we applied CRISPR interference (CRISPRi) technology using deactivated Cas9 (dCas9) to repress genes in <I>C. glutamicum</I>. We then determined the effects of target repression on amino acid titers. Single-guide RNAs directing dCas9 to specific targets reduced expression of <I>pgi</I> and <I>pck</I> up to 98%, and of <I>pyk</I> up to 97%, resulting in titer enhancement ratios of <SMALL>L</SMALL>-lysine and <SMALL>L</SMALL>-glutamate production comparable to levels achieved by gene deletion. This approach for <I>C. glutamicum</I> metabolic engineering, which only requires 3 days, indicates that CRISPRi can be used for quick and efficient metabolic pathway remodeling without the need for gene deletions or mutations and subsequent selection.</P>