A CRISPR/Cas9-based method and primer design tool for seamless genome editing in fission yeast [version 2; peer review: 2 approved]

In the fission yeast Schizosaccharomyces pombe the prevailing approach for gene manipulations is based on homologous recombination of a PCR product that contains genomic target sequences and a selectable marker. The CRISPR/Cas9 system has recently been implemented in fission yeast, which allows for seamless genome editing without integration of a selection marker or leaving any other genomic ‘scars’. The published method involves manual design of the single guide RNA (sgRNA), and digestion of a large plasmid with a problematic restriction enzyme to clone the sgRNA. To increase the efficiency of this approach, we have established and optimized a PCR-based system to clone the sgRNA without restriction enzymes into a plasmid with a dominant natMX6 (nourseothricin) selection marker. We also provide a web-tool, CRISPR4P, to support the design of the sgRNAs and the primers required for the entire process of seamless DNA deletion. Moreover, we report the preparation of G1-synchronized and cryopreserved S. pombe cells, which greatly increases the efficiency and speed for transformations, and may also facilitate standard gene manipulations. Applying this optimized CRISPR/Cas9-based approach, we have successfully deleted over 80 different non-coding RNA genes, which are generally lowly expressed, and have inserted 7 point mutations in 4 different genomic regions

A Highly Instrumented Underground Research Gallery as a Monitoring Concept for Radioactive Waste Cells - Data Measurement Qualification

International audienceThis document presents a dedicated work performed in AndraÕs underground research laboratory (URL) in order to prepare, to test and to qualify the future monitoring system of underground disposal for the French long-lived, intermediate and high level radioactive wastes. Part of the monitoring system qualification process relies on testing sensors on full-scale demonstrators and ensuring that we carefully observe the desired parameters. One of these demonstrators is a concrete liner in a tunnel aiming at support the mechanical pressure of the host rock. A 3.6 meter long section of this gallery has been highly instrumented by various technologies of sensors. This paper describes the monitoring system installed and demonstrates how a numerical model of the demonstrator has been utilized to comfort and validate sensors' measurements

Caspar David Friedrich, the Romantic Hero, and Early German Nationalism

Like many artists, Caspar David Friedrich was affected by the events of his time: the defeat of Prussia by the forces of Napoleon (1806), the following occupation of the German lands (1810), the struggles of the Wars of Liberation (1813-1814) and their disappointing aftermath. He was also greatly influenced by the literary trends of the time, especially the concept of the Romantic hero and the cult of the heroic soldier, ideas that he was exposed to among his circle of friends. The paintings that Friedrich created between 1808 and 1821 display the tensions of the era, emphasizing the feelings of upheaval and desolation experienced in northern Germany and making a statement in favor of a united Fatherland based upon common history and exceptional heroes in a more Romantic fashion than his contemporaries. This paper examines Friedrich’s use of the motif of the hero in his works between 1808 and 1821, including the development of his nationalist ideas and his eventual dismissal of them after the end of the war

\u27In This Dark Hour\u27: Stefan Zweig and Historical Displacement in Brazil, 1941-1942

Stefan Zweig was an Austrian-Jewish author and intellectual who fled Austro-fascism and Nazi Germany, and took his own life in Brazil in early 1942. The resurgence of interest in Zweig’s life in the last few decades has introduced new methods of interpretation of his life as a refugee. But many scholars have not acknowledged Zweig’s relationships he formed with South American intellectuals while in exile there. Instead, the primary focus has been on his identity as a European, and his subsequent suicide. This paper will argue that Zweig’s identity as a refugee included a radical re-interpretation of history and perspective of the world outside of Europe, which had been previously based upon nationalistic and Euro-centric interpretations. Zweig’s exile was one of not only spatial displacement, but was also one of historical displacement, and the physical and political realities in Brazil contributed to this aspect of his life as a refugee

\u27In This Dark Hour\u27: Stefan Zweig and Historical Displacement in Brazil, 1941-1942

Stefan Zweig was an Austrian-Jewish author and intellectual who fled Austro-fascism and Nazi Germany, and took his own life in Brazil in early 1942. The resurgence of interest in Zweig’s life in the last few decades has introduced new methods of interpretation of his life as a refugee. But many scholars have not acknowledged Zweig’s relationships he formed with South American intellectuals while in exile there. Instead, the primary focus has been on his identity as a European, and his subsequent suicide. This paper will argue that Zweig’s identity as a refugee included a radical re-interpretation of history and perspective of the world outside of Europe, which had been previously based upon nationalistic and Euro-centric interpretations. Zweig’s exile was one of not only spatial displacement, but was also one of historical displacement, and the physical and political realities in Brazil contributed to this aspect of his life as a refugee

Cover and Front Matter

Cover, table of content

«Du sublime au ridicule»: Trommel- und Narrenmotiv in Ideen . Buch Le Grand (Heines Reisebilder).

Sin resume

Front matter, vol. 29, issue 2

Editorial board and Advisory Council, masthead, and content

Gene Control during Transcription Elongation

Peer Reviewe