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CRISPR/Cas9 and the microbial cell factory

CRISPR/Cas9, including the derived CRISPRi, has evolved quickly into a popular technology to edit the genomes of prokaryotes and eukaryotes. For constructing microbial cell factories (MCF), it is especially important to regulate metabolic flux toward the target products. To achieve maximal flux channelling toward target products, the regulation of expression of multiple genes is desirable, which is labor intensive and time consuming in most cases. CRISPR/Cas9, especially CRISPRi, allows the design of multiple single strange-guided RNA (sg RNA). These bind to the target genes with different repression intensities with the help of dCas9, and thus regulate metabolic flux. The technology provides a convenient tool to enhance production of various chemicals, regulating cell morphology and changing growth pattern. It is a technology that will revolutionize the metabolic engineering of microbial cells.

  1. Research

    Transcriptional reprogramming in yeast using dCas9 and combinatorial gRNA strategies

    Transcriptional reprogramming is a fundamental process of living cells in order to adapt to environmental and endogenous cues. In order to allow flexible and timely control over gene expression without the int...

    Emil D. Jensen, Raphael Ferreira, Tadas Jakočiūnas, Dushica Arsovska, Jie Zhang, Ling Ding, Justin D. Smith, Florian David, Jens Nielsen, Michael K. Jensen and Jay D. Keasling

    Microbial Cell Factories 2017 16:46

    Published on: 15 March 2017

  2. Research

    CRISPRi-mediated metabolic engineering of E. coli for O-methylated anthocyanin production

    Anthocyanins are a class of brightly colored, glycosylated flavonoid pigments that imbue their flower and fruit host tissues with hues of predominantly red, orange, purple, and blue. Although all anthocyanins ...

    Brady F. Cress, Quentin D. Leitz, Daniel C. Kim, Teresita D. Amore, Jon Y. Suzuki, Robert J. Linhardt and Mattheos A. G. Koffas

    Microbial Cell Factories 2017 16:10

    Published on: 17 January 2017

  3. Research

    CRISPR interference (CRISPRi) for gene regulation and succinate production in cyanobacterium S. elongatus PCC 7942

    Cyanobacterium Synechococcus elongatus PCC 7942 holds promise for biochemical conversion, but gene deletion in PCC 7942 is time-consuming and may be lethal to cells. CRISPR interference (CRISPRi) is an emerging t...

    Chun-Hung Huang, Claire R. Shen, Hung Li, Li-Yu Sung, Meng-Ying Wu and Yu-Chen Hu

    Microbial Cell Factories 2016 15:196

    Published on: 15 November 2016