Asset Details
Do you wish to reserve the book?
In-situ generation of large numbers of genetic combinations for metabolic reprogramming via CRISPR-guided base editing
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
In-situ generation of large numbers of genetic combinations for metabolic reprogramming via CRISPR-guided base editing
Do you wish to request the book?
In-situ generation of large numbers of genetic combinations for metabolic reprogramming via CRISPR-guided base editing
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
In-situ generation of large numbers of genetic combinations for metabolic reprogramming via CRISPR-guided base editing
2021
Overview
Reprogramming complex cellular metabolism requires simultaneous regulation of multigene expression. Ex-situ cloning-based methods are commonly used, but the target gene number and combinatorial library size are severely limited by cloning and transformation efficiencies. In-situ methods such as multiplex automated genome engineering (MAGE) depends on high-efficiency transformation and incorporation of heterologous DNA donors, which are limited to few microorganisms. Here, we describe a Base Editor-Targeted and Template-free Expression Regulation (BETTER) method for simultaneously diversifying multigene expression. BETTER repurposes CRISPR-guided base editors and in-situ generates large numbers of genetic combinations of diverse ribosome binding sites, 5’ untranslated regions, or promoters, without library construction, transformation, and incorporation of DNA donors. We apply BETTER to simultaneously regulate expression of up to ten genes in industrial and model microorganisms
Corynebacterium glutamicum
and
Bacillus subtilis
. Variants with improved xylose catabolism, glycerol catabolism, or lycopene biosynthesis are respectively obtained. This technology will be useful for large-scale fine-tuning of multigene expression in both genetically tractable and intractable microorganisms.
To obtain optimal yield and productivity in bioproduction, expression of pathway genes must be appropriately coordinated. Here, the authors report repurposing of base editors for simultaneous regulation of multiple gene expression and demonstrate its application in industrially important and model microorganisms.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 38/47
/ 42/35
/ 49/31
/ Bacillus subtilis - genetics
/ Bacillus subtilis - metabolism
/ Bacterial Proteins - genetics
/ Bacterial Proteins - metabolism
/ Cloning
/ Corynebacterium glutamicum - genetics
/ Corynebacterium glutamicum - metabolism
/ CRISPR
/ CRISPR-Cas Systems - genetics
/ DNA
/ Genes
/ Genetically engineered microorganisms
/ Genomes
/ Glycerol
/ Humanities and Social Sciences
/ Industrial Microbiology - methods
/ Lycopene
/ Metabolic Engineering - methods
/ Metabolic Networks and Pathways - genetics
/ Science
