Optogenetic methods take advantage of photoswitches to control the activity of cellular proteins. Here, we completed a multi-directional engineering of the fungal photoreceptor Vivid to develop pairs of distinct photoswitches named Magnets. These new photoswitches were engineered to recognize each other based on the electrostatic interactions, thus preventing homodimerization and enhancing light-induced heterodimerization. Furthermore, we tuned the switch-off kinetics by four orders of magnitude and developed several variants, including those with substantially faster kinetics than any of the other conventional dimerization-based blue spectrum photoswitches. We demonstrate the utility of Magnets as powerful tools that can optogenetically manipulate molecular processes in biological systems.

One of the most difficult issues to be solved in genome-wide association studies is to reduce the amount of genomic DNA required for genotyping. Currently available technologies require too large a quantity of genomic DNA to genotype with hundreds or thousands of singlenucleotide polymorphisms (SNPs). To overcome this problem, we combined the Invader assay with multiplex polymerase chain reaction (PCR), carried out in the presence of antibody to Taq polymerase, as well as using a novel 384-well card system that can reduce the required reaction volume. We amplified 100 genomic DNA fragments, each containing one SNP, in a single tube, and analyzed each SNP with the Invader assay. This procedure correctly genotyped 98 of the 100 SNP loci examined in PCR-amplified samples from ten individuals; the genotypes were confirmed by direct sequencing. The reproducibility and universality of the method were confirmed with two additional sets of 100 SNPs. Because we used 40 ng of genomic DNA as a template for multiplex PCR, the amount needed to assay one SNP was only 0.4 ng; therefore, theoretically, more than 200,000 SNPs could be genotyped at once when 100 µg of genomic DNA is available. Our results indicate the feasibility of undertaking genome-wide association studies using blood samples of only 5-10 ml.

Targeted endogenous gene activation is necessary for understanding complex gene networks and has great potential in medical and industrial applications. The CRISPR-Cas system offers simple and powerful tools for this purpose. However, these CRISPR-Cas-based tools for activating user-defined genes are unable to offer precise temporal control of gene expression, despite the fact that many biological phenomena are regulated by highly dynamic patterns of gene expression. Here we created a light-inducible, user-defined, endogenous gene activation system based on CRISPR-Cas9. We demonstrated that this CRISPR-Cas9-based transcription system can allow rapid and reversible targeted gene activation by light. In addition, using this system, we have exemplified photoactivation of multiple user-defined endogenous genes in mammalian cells. The present CRISPR-Cas9-based transcription system offers simple and versatile approaches for precise endogenous gene activation in basic biological research and biotechnology applications.

In recent years, there has been increased interest in topic-focused multi-document summarization. In this task, automatic summaries are produced in response to a specific information request, or topic, stated by the user. The system we have designed to accomplish this task comprises four main components: a generic extractive summarization system, a topic-focusing component, sentence simplification, and lexical expansion of topic words. This paper details each of these components, together with experiments designed to quantify their individual contributions. We include an analysis of our results on two large datasets commonly used to evaluate task-focused summarization, the DUC2005 and DUC2006 datasets, using automatic metrics. Additionally, we include an analysis of our results on the DUC2006 task according to human evaluation metrics. In the human evaluation of system summaries compared to human summaries, i.e., the Pyramid method, our system ranked first out of 22 systems in terms of overall mean Pyramid score; and in the human evaluation of summary responsiveness to the topic, our system ranked third out of 35 systems. AbstractIn recent years, there has been increased interest in topic-focused multi-document summarization. In this task, automatic summaries are produced in response to a specific information request, or topic, stated by the user. The system we have designed to accomplish this task comprises four main components: a generic extractive summarization system, a topic-focusing component, sentence simplification, and lexical expansion of topic words. This paper details each of these components, together with experiments designed to quantify their individual contributions. We include an analysis of our results on two large datasets commonly used to evaluate task-focused summarization, the DUC2005 and DUC2006 datasets, using automatic metrics. Additionally, we include an analysis of our results on the DUC2006 task according to human evaluation metrics. In the human evaluation of system summaries compared to human summaries, i.e., the Pyramid method, our system ranked first out of 22 systems in terms of overall mean Pyramid score; and in the human evaluation of summary responsiveness to the topic, our system ranked third out of 35 systems.