کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6494342 | 44806 | 2015 | 37 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing
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کلمات کلیدی
CRISPRdouble-strand DNAssDNAMMRPEPMEPPAMgRNAG3PMAGEDSBdsDNAORFCRISPR/Cas9 - CRISPR / Cas9Single-strand DNA - DNA تک رشته ایβ-carotene - β-کاروتنclustered regularly interspaced short palindromic repeat - به طور منظم تکرار پیلندرومی کوتاه مدت میان دو طرف تقسیم می شودmismatch repair - تعمیر ناسازگاریguide RNA - راهنمای RNAdouble strand break - شکست دو رشتهphosphoenolpyruvate - فسفوآنولپیرواتopen reading frame - قاب خواندن بازmultiplex automated genome engineering - مهندسی ژنوم اتوماتیک چندگانهprotospacer adjacent motif - موتیف مجاور protospacerGenome editing - ویرایش ژنومglyceraldehyde-3-phosphate - گلیسرالیدید-3-فسفات
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی شیمی
بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing](/preview/png/6494342.png)
چکیده انگلیسی
Engineering cellular metabolism for improved production of valuable chemicals requires extensive modulation of bacterial genome to explore complex genetic spaces. Here, we report the development of a CRISPR-Cas9 based method for iterative genome editing and metabolic engineering of Escherichia coli. This system enables us to introduce various types of genomic modifications with near 100% editing efficiency and to introduce three mutations simultaneously. We also found that cells with intact mismatch repair system had reduced chance to escape CRISPR mediated cleavage and yielded increased editing efficiency. To demonstrate its potential, we used our method to integrate the β-carotene synthetic pathway into the genome and to optimize the methylerythritol-phosphate (MEP) pathway and central metabolic pathways for β-carotene overproduction. We collectively tested 33 genomic modifications and constructed more than 100 genetic variants for combinatorially exploring the metabolic landscape. Our best producer contained15 targeted mutations and produced 2.0 g/L β-carotene in fed-batch fermentation.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Metabolic Engineering - Volume 31, September 2015, Pages 13-21
Journal: Metabolic Engineering - Volume 31, September 2015, Pages 13-21
نویسندگان
Yifan Li, Zhenquan Lin, Can Huang, Yan Zhang, Zhiwen Wang, Ya-jie Tang, Tao Chen, Xueming Zhao,