Institution
University of Stuttgart
Education•Stuttgart, Germany•
About: University of Stuttgart is a education organization based out in Stuttgart, Germany. It is known for research contribution in the topics: Laser & Finite element method. The organization has 27715 authors who have published 56370 publications receiving 1363382 citations. The organization is also known as: Universität Stuttgart.
Papers published on a yearly basis
Papers
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TL;DR: In this article, a high resolution (2 μm) acoustic step-wave probing technique has been developed to investigate the inhomogeneous distribution of the piezoelectric response of incompletely corona poled 85 μm thick PVDF films.
220 citations
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TL;DR: The single-plasmid system constructed for the genome editing of B. subtilis overcomes the problems of counterselection methods and may be adapted for use in other firmicutes.
Abstract: The clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) systems are adaptive immune systems of bacteria. A type II CRISPR-Cas9 system from Streptococcus pyogenes has recently been developed into a genome engineering tool for prokaryotes and eukaryotes. Here, we present a single-plasmid system which allows efficient genome editing of Bacillus subtilis. The plasmid pJOE8999 is a shuttle vector that has a pUC minimal origin of replication for Escherichia coli, the temperature-sensitive replication origin of plasmid pE194ts for B. subtilis, and a kanamycin resistance gene working in both organisms. For genome editing, it carries the cas9 gene under the control of the B. subtilis mannose-inducible promoter PmanP and a single guide RNA (sgRNA)-encoding sequence transcribed via a strong promoter. This sgRNA guides the Cas9 nuclease to its target. The 20-nucleotide spacer sequence at the 5′ end of the sgRNA sequence, responsible for target specificity, is located between BsaI sites. Thus, the target specificity is altered by changing the spacer sequences via oligonucleotides fitted between the BsaI sites. Cas9 in complex with the sgRNA induces double-strand breaks (DSBs) at its target site. Repair of the DSBs and the required modification of the genome are achieved by adding homology templates, usually two PCR fragments obtained from both sides of the target sequence. Two adjacent SfiI sites enable the ordered integration of these homology templates into the vector. The function of the CRISPR-Cas9 vector was demonstrated by introducing two large deletions in the B. subtilis chromosome and by repair of the trpC2 mutation of B. subtilis 168.
IMPORTANCE In prokaryotes, most methods used for scarless genome engineering are based on selection-counterselection systems. The disadvantages are often the lack of a suitable counterselection marker, the toxicity of the compounds needed for counterselection, and the requirement of certain mutations in the target strain. CRISPR-Cas systems were recently developed as important tools for genome editing. The single-plasmid system constructed for the genome editing of B. subtilis overcomes the problems of counterselection methods. It allows deletions and introduction of point mutations. It is easy to handle and very efficient, and it may be adapted for use in other firmicutes.
220 citations
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220 citations
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TL;DR: In this paper, a semi-quantitative nonlinear field theory of the brain under realistic anatomical connectivity conditions describing the interaction between functional units within the brain is presented, which is derived from the quasi-microscopic conversion properties of neural populations occurring at synapses and somas.
219 citations
Authors
Showing all 28043 results
Name | H-index | Papers | Citations |
---|---|---|---|
Yi Chen | 217 | 4342 | 293080 |
Robert J. Lefkowitz | 214 | 860 | 147995 |
Michael Kramer | 167 | 1713 | 127224 |
Andrew G. Clark | 140 | 823 | 123333 |
Stephen D. Walter | 112 | 513 | 57012 |
Fedor Jelezko | 103 | 413 | 42616 |
Ulrich Gösele | 102 | 603 | 46223 |
Dirk Helbing | 101 | 642 | 56810 |
Ioan Pop | 101 | 1370 | 47540 |
Niyazi Serdar Sariciftci | 99 | 591 | 54055 |
Matthias Komm | 99 | 832 | 43275 |
Hans-Joachim Werner | 98 | 317 | 48508 |
Richard R. Ernst | 96 | 352 | 53100 |
Xiaoming Sun | 96 | 382 | 47153 |
Feng Chen | 95 | 2138 | 53881 |