Abstract：
:Multiplexed gene expression optimization via modulation of gene translation efficiency through ribosome binding site (RBS) engineering is a valuable approach for optimizing artificial properties in bacteria, ranging from genetic circuits to production pathways. Established algorithms design smart RBS-libraries based on a single partially-degenerate sequence that efficiently samples the entire space of translation initiation rates. However, the sequence space that is accessible when integrating the library by CRISPR/Cas9-based genome editing is severely restricted by DNA mismatch repair (MMR) systems. MMR efficiency depends on the type and length of the mismatch and thus effectively removes potential library members from the pool. Rather than working in MMR-deficient strains, which accumulate off-target mutations, or depending on temporary MMR inactivation, which requires additional steps, we eliminate this limitation by developing a pre-selection rule of genome-library-optimized-sequences (GLOS) that enables introducing large functional diversity into MMR-proficient strains with sequences that are no longer subject to MMR-processing. We implement several GLOS-libraries in Escherichia coli and show that GLOS-libraries indeed retain diversity during genome editing and that such libraries can be used in complex genome editing operations such as concomitant deletions. We argue that this approach allows for stable and efficient fine tuning of chromosomal functions with minimal effort.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Oesterle S,Gerngross D,Schmitt S,Roberts TM,Panke Sdoi
10.1038/s41598-017-12395-3subject
Has Abstractpub_date
2017-09-26 00:00:00pages
12327issue
1issn
2045-2322pii
10.1038/s41598-017-12395-3journal_volume
7pub_type
杂志文章abstract：:Archeological researches have proposed arguments for human mobility and long-distance trading over the Eurasia before the Silk Roads. Here we utilize biologically available strontium isotope analysis to assess the extent of pre-Silk Road population movements and cultural communications across the Asian interior. From ...
journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2016-10-20 00:00:00
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pub_type： 杂志文章
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pub_type： 杂志文章
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pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2017-01-23 00:00:00
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pub_type： 杂志文章
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pub_type： 已发布勘误
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更新日期：2020-07-29 00:00:00
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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更新日期：2017-01-13 00:00:00
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pub_type： 杂志文章
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journal_title：Scientific reports
pub_type： 杂志文章
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journal_title：Scientific reports
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