CRISPR/Cas9-based genome editing has been one of the major achievements of molecular biology,allowing the targeted engineering of a wide range of genomes.The system originally evolved in prokaryotes as an adaptive imm...CRISPR/Cas9-based genome editing has been one of the major achievements of molecular biology,allowing the targeted engineering of a wide range of genomes.The system originally evolved in prokaryotes as an adaptive immune system against bacteriophage infections.It now sees widespread application in genome engineering workflows,especially using the Streptococcus pyogenes endonuclease Cas9.To utilize Cas9,so-called single guide RNAs(sgRNAs)need to be designed for each target gene.While there are many tools available to design sgRNAs for the popular model organisms,only few tools that allow designing sgRNAs for non-model organisms exist.Here,we present CRISPy-web(http://crispy.secondarymetabolites.org/),an easy to use web tool based on CRISPy to design sgRNAs for any userprovided microbial genome.CRISPy-web allows researchers to interactively select a region of their genome of interest to scan for possible sgRNAs.After checks for potential off-target matches,the resulting sgRNA sequences are displayed graphically and can be exported to text files.All steps and information are accessible from a web browser without the requirement to install and use command line scripts.展开更多
To accelerate the shift to bio-based production and overcome complicated functional implementation of natural and artificial biosynthetic pathways to industry relevant organisms,development of new,versatile,bio-based ...To accelerate the shift to bio-based production and overcome complicated functional implementation of natural and artificial biosynthetic pathways to industry relevant organisms,development of new,versatile,bio-based production platforms is required.Here we present a novel yeast-based platform for biosynthesis of bacterial aromatic polyketides.The platform is based on a synthetic polyketide synthase system enabling a first demonstration of bacterial aromatic polyketide biosynthesis in a eukaryotic host.展开更多
Energy crops play a vital role in meeting future energy and chemical demands while addressing climate change.However,the idealization of lowcarbon workflows and careful consideration of cost-benefit equations are cruc...Energy crops play a vital role in meeting future energy and chemical demands while addressing climate change.However,the idealization of lowcarbon workflows and careful consideration of cost-benefit equations are crucial for their more sustainable implementation.Here,we propose tobacco as a promising energy crop because of its exceptional water solubility,mainly attributed to a high proportion of water-soluble carbohydrates and nitrogen,less lignocellulose,and the presence of acids.We then designed a strategy that maximizes biomass conversion into bio-based products while minimizing energy and material inputs.By autoclaving tobacco leaves in water,we obtained a nutrient-rich medium capable of supporting the growth of microorganisms and the production of bioproducts without the need for extensive pretreatment,hydrolysis,or additional supplements.Additionally,cultivating tobacco on barren lands can generate sufficient biomass to produce approximately 573 billion gallons of ethanol per year.This approach also leads to a reduction of greenhouse gas emissions by approximately 76%compared to traditional corn stover during biorefinery processes.Therefore,our study presents a novel and direct strategy that could significantly contribute to the goal of reducing carbon emissions and global sustainable development compared to traditional methods.展开更多
文摘CRISPR/Cas9-based genome editing has been one of the major achievements of molecular biology,allowing the targeted engineering of a wide range of genomes.The system originally evolved in prokaryotes as an adaptive immune system against bacteriophage infections.It now sees widespread application in genome engineering workflows,especially using the Streptococcus pyogenes endonuclease Cas9.To utilize Cas9,so-called single guide RNAs(sgRNAs)need to be designed for each target gene.While there are many tools available to design sgRNAs for the popular model organisms,only few tools that allow designing sgRNAs for non-model organisms exist.Here,we present CRISPy-web(http://crispy.secondarymetabolites.org/),an easy to use web tool based on CRISPy to design sgRNAs for any userprovided microbial genome.CRISPy-web allows researchers to interactively select a region of their genome of interest to scan for possible sgRNAs.After checks for potential off-target matches,the resulting sgRNA sequences are displayed graphically and can be exported to text files.All steps and information are accessible from a web browser without the requirement to install and use command line scripts.
基金This work was funded by grants from the Novo Nordisk Foundation[NNF10CC1016517],[NNF15OC0016626]and is part of the U.S.Department of Energy Joint BioEnergy Institute supported by the U.S.Department of Energy,Office of Science,Office of Biological and Environmental Research,through Contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the U.S.Department of Energy.
文摘To accelerate the shift to bio-based production and overcome complicated functional implementation of natural and artificial biosynthetic pathways to industry relevant organisms,development of new,versatile,bio-based production platforms is required.Here we present a novel yeast-based platform for biosynthesis of bacterial aromatic polyketides.The platform is based on a synthetic polyketide synthase system enabling a first demonstration of bacterial aromatic polyketide biosynthesis in a eukaryotic host.
基金financial supports from the Shandong Provincial Natural Science Foundation(numbers ZR2020JQ11 and ZR2023QC246)the Qingdao New Energy Shandong Laboratory Open Project(number QNESL OP202308)+4 种基金the National Natural Science Foundation of China(numbers NSF32170084 and NSF32170387)Young Taishan Scholars(number TSQN201909159)the Novo Nordisk Fonden(number NNF20CC0035580)the Agricultural Science and Technology Innovation Program(number ASTIP-TRIC02)the Youth Innovation Promotion Association,CAS(number Y2021063).
文摘Energy crops play a vital role in meeting future energy and chemical demands while addressing climate change.However,the idealization of lowcarbon workflows and careful consideration of cost-benefit equations are crucial for their more sustainable implementation.Here,we propose tobacco as a promising energy crop because of its exceptional water solubility,mainly attributed to a high proportion of water-soluble carbohydrates and nitrogen,less lignocellulose,and the presence of acids.We then designed a strategy that maximizes biomass conversion into bio-based products while minimizing energy and material inputs.By autoclaving tobacco leaves in water,we obtained a nutrient-rich medium capable of supporting the growth of microorganisms and the production of bioproducts without the need for extensive pretreatment,hydrolysis,or additional supplements.Additionally,cultivating tobacco on barren lands can generate sufficient biomass to produce approximately 573 billion gallons of ethanol per year.This approach also leads to a reduction of greenhouse gas emissions by approximately 76%compared to traditional corn stover during biorefinery processes.Therefore,our study presents a novel and direct strategy that could significantly contribute to the goal of reducing carbon emissions and global sustainable development compared to traditional methods.
