Thioester Electrophiles as a New Class of SHAPE Probes

RNA is a multifunctional molecule capable of regulating gene expression, in large part because it can form a variety of RNA secondary and tertiary structures. The emergence of RNA viruses like SARS-CoV-2 emphasizes the need to accelerate our understanding of how viral RNA structure dictates its function. One approach to map RNA secondary structure, called Selective 2'-OH Acylation Analyzed by Primer Extension (SHAPE), utilizes select electrophiles that unbiasedly modifies the 2'-hydroxyl of riboses in unpaired nucleotides, forming adducts that are detected through a variety of sequencing methods. While SHAPE is widely utilized, most existing SHAPE reagents suffer from several drawbacks: 1) poor water solubility;2) limited commercial availability;and 3) they function optimally when freshly synthesized, requiring synthetic organic expertise. To overcome these obstacles, our goal is to develop a userfriendly SHAPE reagent kit that provides highly reactive, soluble SHAPE reagents capable of probing RNA structure in vitro as well as in vivo. We present our investigations on developing thioester electrophiles as a new class of SHAPE reagents. Our reagent is prepared by mixing two stable components to generate the reactive thioester electrophile in situ. We report our preliminary results in model systems and the scope to expand the library of our reagents.Copyright © 2023 The American Society for Biochemistry and Molecular Biology, Inc.

COVID-19 infection rate following COVID-19 vaccination among healthcare professionals at a tertiary care public hospital in city of Mumbai

Background: India launched COVID-19 mass Vaccination campaign after regulatory approval of Covishield & Covaxin vaccines. However, inspite of Vaccination, cases of COVID-19 infection are being reported. Hence, this study is aimed to assess the COVID-19 infection and/or re-infection rate, including breakthrough infections, following vaccination among Health Care Workers at a Tertiary Care Dedicated COVID Hospital. This questionnaire-based survey was initiated following Institutional Ethics Committee approval. We also looked at reasons for Vaccine hesitancy and occurrence of adverse reactions following vaccination, their management and duration amongst the Healthcare Workers. Results: Of 564 Healthcare Workers (HCWs) who consented to participate, only 503 filled in the questionnaire completely. Majority of the HCWs received Covishield vaccine (78.56%). The infection rate postvaccination was 8.28% (with a median of 22 days and IQR of 8-43 days). This infection rate was significantly higher in those who were not vaccinated as compared to the Vaccinated HCWs (OR = 0.10, 95 CI% = 0.05-0.22, p <0.0001). Breakthrough infection rate was 2.42. Although 58.39% of the participants suffered adverse reactions after vaccination, like myalgia, Fever, Headache, these were mild in nature lasting for an average of 3-4 days. The vaccine hesitancy rate at our hospital HCWs was 6.36%, the main issue being the concerns regarding safety and effectiveness of the vaccines against the COVID-19 infection. Discussion/Conclusion: The infection and Breakthrough infection rates in our study were low and severity of COVID infection post vaccination was mild, not requiring hospitalisation.