Advances and applications of CRISPR/Cas-mediated interference in Escherichia coli

Non-native Pathway Engineering with CRISPRi for Carbon Dioxide Assimilation and Valued 5-Aminolevulinic Acid Synthesis in Escherichia coli Nissle

“…Moreover, to enhance flux toward the RuBisCO pathway, zwf , pfk A, and pfk B genes were knocked down, avoiding carbon loss (i.e., CO 2 release) (Figure A). , CRISPRi-based gene regulation was chosen due to its high efficiency and flexibility in multiple targeting specific loci in the genome. Additionally, CRISPRi requires only one protein, dCas9, to interfere with gene transcription by allosterically binding to the target gene . The effect of CRISPRi design was evaluated by coexpressing the sgRNA plasmid with the CO 2 -fixing genes in glucose- and xylose-based mediums.…”

“…Additionally, CRISPRi requires only one protein, dCas9, to interfere with gene transcription by allosterically binding to the target gene. 32 The effect of CRISPRi design was evaluated by coexpressing the sgRNA plasmid with the CO 2 -fixing genes in glucose- and xylose-based mediums. In the glucose medium, strains imposed on the CRISPRi system showed strong cell burden and limited growth.…”