RNA Editing by Adenosine Deaminase Acting on RNA 1 Under Different Metabolic Conditions - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook RNA Editing by Adenosine Deaminase Acting on RNA 1 Under Different Metabolic Conditions write by Kira Schmiedeknecht. This book was released on 2024. RNA Editing by Adenosine Deaminase Acting on RNA 1 Under Different Metabolic Conditions available in PDF, EPUB and Kindle.
Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing
Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing write by Charles E. Samuel. This book was released on 2011-11-06. Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing available in PDF, EPUB and Kindle. “The objective of this CTMI volume is to provide readers with a foundation for understanding what ADARs are and how they act to affect gene expression and function. It is becoming increasingly apparent that ADARs may possess roles not only as enzymes that deaminate adenosine to produce inosine in RNA substrates with double-stranded character, but also as proteins independent of their catalytic property. Because A-to-I editing may affect base-pairing and RNA structure, processes including translation, splicing, RNA replication, and miR and siRNA silencing may be affected. Future studies of ADARs no doubt will provide us with additional surprises and new insights into the modulation of biological processes by the ADAR family of proteins.”
Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing
Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing write by Charles E. Samuel. This book was released on 2011-11-09. Adenosine Deaminases Acting on RNA (ADARs) and A-to-I Editing available in PDF, EPUB and Kindle. “The objective of this CTMI volume is to provide readers with a foundation for understanding what ADARs are and how they act to affect gene expression and function. It is becoming increasingly apparent that ADARs may possess roles not only as enzymes that deaminate adenosine to produce inosine in RNA substrates with double-stranded character, but also as proteins independent of their catalytic property. Because A-to-I editing may affect base-pairing and RNA structure, processes including translation, splicing, RNA replication, and miR and siRNA silencing may be affected. Future studies of ADARs no doubt will provide us with additional surprises and new insights into the modulation of biological processes by the ADAR family of proteins.”
Structural Engineering of Adenosine Deaminases Acting on RNA with Chemically Modified Guide RNAs for Site-directed RNA Editing
Structural Engineering of Adenosine Deaminases Acting on RNA with Chemically Modified Guide RNAs for Site-directed RNA Editing - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook Structural Engineering of Adenosine Deaminases Acting on RNA with Chemically Modified Guide RNAs for Site-directed RNA Editing write by Leanna Rose Monteleone. This book was released on 2020. Structural Engineering of Adenosine Deaminases Acting on RNA with Chemically Modified Guide RNAs for Site-directed RNA Editing available in PDF, EPUB and Kindle. RNA editing is defined as the insertion, deletion, or modification of a nucleotide that changes the information content of a sequence. Adenosine deaminases acting on RNA (ADARs) can deaminate an adenosine (A) in duplex RNA to inosine (I). Cellular machinery interprets inosine as guanosine, which can result in various consequences on RNA function. A-to-I editing can alter microRNA sequences, redirect splicing, and change secondary structure. More dramatically A-to-I editing can result in a recoding event, thereby changing the amino acid at a specific position. In recent years, there has been rapidly growing interest in engineering ADARs or directing endogenous ADARs to specific G-to-A mutations linked to various diseases. The contents of this dissertation details the progress we have made, with the help of various collaborations, to use ADARs for site- directed RNA editing. In chapter 1, I review various types of RNA editing with a great focus on adenosine deamination. I emphasize ADARs biological function, substrate specificity, and the roles ADARs have in various diseases. I further discuss the structural data that is known for ADAR2 and how this knowledge has led to a better understanding of using ADARs for site-directed RNA editing. In chapter 2, I discuss the previous approaches used for site-ivdirected RNA editing with ADAR and the challenge of overcoming off-target reactions. To overcome off-target reactions, I have designed an orthogonal editing system utilizing a bump-hole strategy to prevent off-target edits. I have shown that combining bulky ADAR mutants with a chemically modified guide RNA (gRNA) achieves site-selective editing with reduced off-target edits both in vitro and in cellular assays. In chapter 3, I focus on our collaboration with Prof. Gail Mandel's laboratory at Oregon Health and Science University to study a disease-causing mutation linked to Rett Syndrome. In this approach, we have focused on rationally designing chemically modified gRNAs that could potentially recruit endogenous wild type ADARs. Our rational design utilizes the crystallography of ADAR2 constructs bound to double stranded RNA (dsRNA) that were solved by our collaborators in Prof. Andrew Fisher's laboratory. In chapter 4, I deviate from using ADARs for site-directed RNA editing to elucidate the biological role of ADAR3. ADAR3 is catalytically inactive and is exclusively located in the brain. To further understand the role of ADAR3, five mutations were incorporated to engineer an active ADAR3 (ADAR3 M3). From here, we propose that ADAR3 not only acts as a negative regulator of ADAR1 and ADAR2, but also as a direct regulator in stabilizing specific transcripts. With an active ADAR3, future studies can be done to use ADAR3 M3 or another version of an active ADAR3 for site-directed RNA editing.
RNA Recognition by Adenosine Deaminases Acting on RNA
RNA Recognition by Adenosine Deaminases Acting on RNA - read free eBook in online reader or directly download on the web page. Select files or add your book in reader. Download and read online ebook RNA Recognition by Adenosine Deaminases Acting on RNA write by Yuru Wang. This book was released on 2017. RNA Recognition by Adenosine Deaminases Acting on RNA available in PDF, EPUB and Kindle. Adenosine deaminases acting on RNA (ADAR) catalyze adenosine to inosine changes in double stranded RNAs, a type of post-transcriptional modification that can change the codon meaning and contribute to protein diversity in higher organisms. This modification is also known to regulate the fate of the RNA, including its expression, turnover, involvement in RNA interference and so forth. Three types of ADARs have been found in mammals, with ADAR1 and ADAR2 being catalytically active whereas ADAR3 being considered catalytically inactive. Malfunctions of ADARs have been correlated with various human diseases, including cancer. The Beal lab over the years has devoted extensive efforts in elucidating how ADARs recognize RNA substrates, and understanding the mechanism behind the RNA recognition difference between ADAR1 and ADAR2. These efforts not only advance our understanding of how these enzymes function, but also pave the way for future development of ADAR specific inhibitors of therapeutic significance. This thesis is a continuation of these efforts contributing to our understanding of how these fascinating enzymes function and providing new tools for future studies of them. Chapter 1 is an introduction of background knowledge about A-to-I RNA editing and ADAR. Chapter 2 introduced a new phenotypic reporter system that utilizes an RNA substrate efficiently processed by both ADAR1 and ADAR2 catalytic domains (ADAR-D) and a study utilizing this reporter to probe the RNA recognition by the base flipping residue in ADAR1. On the basis of this reporter system, in Chapter 3, a fluorescent reporter assay was developed to achieve high-throughput and quantitative evaluation of ADAR editing activity never achieved by other assays before, and a method called Sat-FACS-seq was introduced which provides information-rich landscape of sequence requirement across any region in ADARs. Applying this method to the 5’ binding loop of ADAR2, a novel insight into how this loop recognizes RNA was obtained. Chapter 4 detailed a study on the RNA secondary structural features that could distinguish ADAR1-D editing from ADAR2-D editing. Experimental evidence was shown, for the first time, to prove that the 5’ binding loops contribute to the site selectivity difference between ADAR1 and ADAR2, probably through differential recognition of RNA structure in the region 5’ from the editing site. Lastly, in Chapter 5, an effort to evolve the inactive ADAR3 into an active deaminase was described. Our success in turning ADAR3 into an active deaminase not only provides structural explanation of why wild-type ADAR3 is catalytically inactive, but also advances our knowledge of important residues required for proper ADAR function other than the ones traditionally appreciated. Moreover, the active ADAR3 mutant obtained was introduced with a minimal number of mutations (five), none of which was located in the RNA binding domains or on the primary RNA recognition surfaces. Thus, the mutant would be of great value for identifying the cellular binding targets of ADAR3 in vivo, which is important for understanding its biological function.
