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      Raman Spectroscopy for Chemical Biology Research

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          Abstract

          In chemical biology research, various fluorescent probes have been developed and used to visualize target proteins or molecules in living cells and tissues, yet there are limitations to this technology, such as the limited number of colors that can be detected simultaneously. Recently, Raman spectroscopy has been applied in chemical biology to overcome such limitations. Raman spectroscopy detects the molecular vibrations reflecting the structures and chemical conditions of molecules in a sample and was originally used to directly visualize the chemical responses of endogenous molecules. However, our initial research to develop “Raman tags” opens a new avenue for the application of Raman spectroscopy in chemical biology. In this Perspective, we first introduce the label-free Raman imaging of biomolecules, illustrating the biological applications of Raman spectroscopy. Next, we highlight the application of Raman imaging of small molecules using Raman tags for chemical biology research. Finally, we discuss the development and potential of Raman probes, which represent the next-generation probes in chemical biology.

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          Most cited references167

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          A chemical method for fast and sensitive detection of DNA synthesis in vivo.

          We have developed a method to detect DNA synthesis in proliferating cells, based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU) and its subsequent detection by a fluorescent azide through a Cu(I)-catalyzed [3 + 2] cycloaddition reaction ("click" chemistry). Detection of the EdU label is highly sensitive and can be accomplished in minutes. The small size of the fluorescent azides used for detection results in a high degree of specimen penetration, allowing the staining of whole-mount preparations of large tissue and organ explants. In contrast to BrdU, the method does not require sample fixation or DNA denaturation and permits good structural preservation. We demonstrate the use of the method in cultured cells and in the intestine and brain of whole animals.
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            The CRISPR tool kit for genome editing and beyond

            CRISPR is becoming an indispensable tool in biological research. Once known as the bacterial immune system against invading viruses, the programmable capacity of the Cas9 enzyme is now revolutionizing diverse fields of medical research, biotechnology, and agriculture. CRISPR-Cas9 is no longer just a gene-editing tool; the application areas of catalytically impaired inactive Cas9, including gene regulation, epigenetic editing, chromatin engineering, and imaging, now exceed the gene-editing functionality of WT Cas9. Here, we will present a brief history of gene-editing tools and describe the wide range of CRISPR-based genome-targeting tools. We will conclude with future directions and the broader impact of CRISPR technologies.
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              Prostaglandins and leukotrienes: advances in eicosanoid biology.

              C D Funk (2001)
              Prostaglandins and leukotrienes are potent eicosanoid lipid mediators derived from phospholipase-released arachidonic acid that are involved in numerous homeostatic biological functions and inflammation. They are generated by cyclooxygenase isozymes and 5-lipoxygenase, respectively, and their biosynthesis and actions are blocked by clinically relevant nonsteroidal anti-inflammatory drugs, the newer generation coxibs (selective inhibitors of cyclooxygenase-2), and leukotriene modifiers. The prime mode of prostaglandin and leukotriene action is through specific G protein-coupled receptors, many of which have been cloned recently, thus enabling specific receptor agonist and antagonist development. Important insights into the mechanisms of inflammatory responses, pain, and fever have been gleaned from our current understanding of eicosanoid biology.
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                Author and article information

                Journal
                J Am Chem Soc
                J Am Chem Soc
                ja
                jacsat
                Journal of the American Chemical Society
                American Chemical Society
                0002-7863
                1520-5126
                10 October 2022
                02 November 2022
                10 October 2023
                : 144
                : 43
                : 19651-19667
                Affiliations
                []Synthetic Organic Chemistry Laboratory, RIKEN Cluster for Pioneering Research , 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
                []Catalysis and Integrated Research Group, RIKEN Center for Sustainable Resource Science , 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
                [§ ]Department of Applied Physics, Osaka University , 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan
                []Institute for Open and Transdisciplinary Research Initiatives, Osaka University , Suita, Osaka 565-0871, Japan
                [# ]AIST-Osaka University Advanced Photonics and Biosensing Open Innovation Laboratory, National Institute of Advanced Industrial Science and Technology (AIST) , Suita, Osaka 565-0871, Japan
                Author notes
                Author information
                https://orcid.org/0000-0001-6008-2915
                https://orcid.org/0000-0002-2284-375X
                https://orcid.org/0000-0002-1344-364X
                Article
                10.1021/jacs.2c05359
                9635364
                36216344
                e6b230a1-3266-495e-878a-c89cd5548c53
                © 2022 The Authors. Published by American Chemical Society

                Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works ( https://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 21 May 2022
                Funding
                Funded by: Japan Agency for Medical Research and Development, doi 10.13039/100009619;
                Award ID: JP17gm0710004
                Funded by: Japan Science and Technology Agency, doi 10.13039/501100002241;
                Award ID: JPMJCR1925
                Funded by: Japan Science and Technology Agency, doi 10.13039/501100002241;
                Award ID: ERATO Live Cell Chemistry Project
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                ja2c05359
                ja2c05359

                Chemistry
                Chemistry

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