Metabolic engineering of Escherichia coli using CRISPR–Cas9 meditated genome editing

• Published in: Metabolic engineering Vol. 31; pp. 13 - 21
• Main Authors: Li, Yifan, Lin, Zhenquan, Huang, Can, Zhang, Yan, Wang, Zhiwen, Tang, Ya-jie, Chen, Tao, Zhao, Xueming
• Format: Journal Article
• Language: English
• Published: Belgium Elsevier Inc 01.09.2015
• Subjects: base pair mismatch; batch fermentation; beta Carotene - biosynthesis; beta-carotene; biochemical pathways; Clustered Regularly Interspaced Short Palindromic Repeats - genetics; Combinatorial metabolic engineering; CRISPR-Cas Systems; CRISPR/Cas9; DNA Mismatch Repair; DNA repair; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; genetic variation; genome; Genome editing; Genome, Bacterial; Metabolic Engineering; RNA Editing - genetics; β-Carotene; CRISPR; ORF; TRMR; CRISPR/Cas9; G3P; gRNA; Combinatorial metabolic engineering; β-Carotene; MAGE; DSB; MMR; MEP; PEP; Genome editing; PAM; ssDNA; dsDNA

Engineering cellular metabolism for improved production of valuable chemicals requires extensive modulation of bacterial genome to explore complex genetic spaces. Here, we report the development of a CRISPR–Cas9 based method for iterative genome editing and metabolic engineering of Escherichia coli....