If a person(known as the agent) acts without authority,or exceeds his authority,his actions cannot normally bind the person(known as the principal) on whose behalf he purports to act.However,subsequently,ratification ...If a person(known as the agent) acts without authority,or exceeds his authority,his actions cannot normally bind the person(known as the principal) on whose behalf he purports to act.However,subsequently,ratification allows the principal to adopt and endorse the agent’s acts.A remarkable feature of ratification is that it is retroactive in effect thereby becoming operative from the moment of the agent’s contracting with the third party rather than the moment of the principal’s ratification.展开更多
The bacterium Escherichia coli(E.coli)is one of the most widely used chassis microbes employed for the biosynthesis of numerous valuable chemical compounds.In the past decade,the metabolic engineering of E.coli has un...The bacterium Escherichia coli(E.coli)is one of the most widely used chassis microbes employed for the biosynthesis of numerous valuable chemical compounds.In the past decade,the metabolic engineering of E.coli has undergone significant advances,although further productivity improvements will require extensive genome modification,multi-dimensional regulation,and multiple metabolic-pathway coordination.In this context,clustered regularly interspaced short palindromic repeats(CRISPR),along with CRISPR-associated protein(Cas)and its inactive variant(dCas),have emerged as notable recombination and transcriptional regulation tools that are particularly useful for multiplex metabolic engineering in E.coli.In this review,we briefly describe the CRISPR/Cas9 technology in E.coli,and then summarize the recent advances in CRISPR/dCas9 interference(CRISPRi)systems in E.coli,particularly the strategies designed to effectively regulate gene repression and overcome retroactivity during multiplexing.Moreover,we discuss recent applications of the CRISPRi system for enhancing metabolite production in E.coli,and finally highlight the major challenges and future perspectives of this technology.展开更多
文摘If a person(known as the agent) acts without authority,or exceeds his authority,his actions cannot normally bind the person(known as the principal) on whose behalf he purports to act.However,subsequently,ratification allows the principal to adopt and endorse the agent’s acts.A remarkable feature of ratification is that it is retroactive in effect thereby becoming operative from the moment of the agent’s contracting with the third party rather than the moment of the principal’s ratification.
基金supported by the Agency for Science,Technology and Research(A∗STAR)under Central Research Fund(Applied/Translational Research),AME Young Individual.Research Grant:A2084c0064(2019).
文摘The bacterium Escherichia coli(E.coli)is one of the most widely used chassis microbes employed for the biosynthesis of numerous valuable chemical compounds.In the past decade,the metabolic engineering of E.coli has undergone significant advances,although further productivity improvements will require extensive genome modification,multi-dimensional regulation,and multiple metabolic-pathway coordination.In this context,clustered regularly interspaced short palindromic repeats(CRISPR),along with CRISPR-associated protein(Cas)and its inactive variant(dCas),have emerged as notable recombination and transcriptional regulation tools that are particularly useful for multiplex metabolic engineering in E.coli.In this review,we briefly describe the CRISPR/Cas9 technology in E.coli,and then summarize the recent advances in CRISPR/dCas9 interference(CRISPRi)systems in E.coli,particularly the strategies designed to effectively regulate gene repression and overcome retroactivity during multiplexing.Moreover,we discuss recent applications of the CRISPRi system for enhancing metabolite production in E.coli,and finally highlight the major challenges and future perspectives of this technology.
