Yeast
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Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology is an important tool for genome editing because the Cas9 endonuclease can induce targeted DNA double-strand breaks. Targeting of the DNA break is typically controlled by a single-guide RNA (sgRNA), a chimeric RNA containing a structural segment important for Cas9 binding and a 20mer guide sequence that hybridizes to the genomic DNA target. Previous studies have demonstrated that CRISPR-Cas9 technology can be used for efficient, marker-free genome editing in Saccharomyces cerevisiae. ⋯ Here we describe a flexible set of vectors based on this design for cloning and expressing sgRNAs (and Cas9) in yeast using different selectable markers. We anticipate that the Cas9-sgRNA expression vector with the URA3 selectable marker (pML104) will be particularly useful for genome editing in yeast, since the Cas9 machinery can be easily removed by counter-selection using 5-fluoro-orotic acid (5-FOA) following successful genome editing. The availability of new vectors that simplify and streamline the technical steps required for guide sequence cloning should help accelerate the use of CRISPR-Cas9 technology in yeast genome editing.
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While yeast apoptosis was still a controversial issue less than 10 years ago, the efforts of many groups have revealed cell death mechanisms that resemble, in many aspects, those described for mammalian apoptosis. Here, we provide an overview of new insights on yeast apoptosis and the link with lifespan of yeast cells, based on data presented at the 6th International Meeting of Yeast Apoptosis (IMYA). Together, these data demonstrate the power and advantages of the yeast system to uncover novel cellular factors governing life and death, placing yeast at the forefront of apoptosis research.
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UK CropNet currently provides a range of databases (and database-mining tools) to the plant community that are all freely accessible through our website (http://ukcrop.net/). Recent upgrades have meant that we can now expand the range of available facilities (e.g. addition of new databases) whilst also strengthening and improving access to existing services (e.g. providing a BLAST search facility against sequences in our databases). This article will briefly outline these and other new developments in our service.