Background: Modified nucleotides are increasingly being utilized in the de novo selection of aptamers for enhancing their drug-like character and abolishing the need for time consuming trial-and-error based post-selection modifications. Locked nucleic acid (LNA) is one of the most prominent and successful nucleic acid analogues because of its remarkable properties, and widely explored as building blocks in therapeutic oligonucleotides. Evolution of LNA-modified RNA aptamers requires an efficient reverse transcription method for PCR enrichment of the selected RNA aptamer candidates. Establishing this key step is a pre-requisite for performing LNA-modified RNA aptamer selection.
Methodology: In this study three different reverse transcriptases were investigated towards the enzymatic recognition of LNA nucleotides. Both incorporation as well as reading capabilities of the LNA nucleotides was investigated to fully understand the limitations of the enzymatic recognition.
Conclusions: We found that SuperScript® III Reverse Transcriptase is an efficient enzyme for the recognition of LNA nucleotides, making it a prime candidate to be used in de novo selection of LNA containing RNA aptamers.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3338489 | PMC |
| http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0035990 | PLOS |
Publication Analysis
Top Keywords
Similar Publications
Analyzing bacterial networks and interactions in skin and gills of Sparus aurata with microalgae-based additive feeding.
Sci Rep
December 2024
Department of Microbiology, Faculty of Sciences, CEI·MAR-International Campus of Excellence in Marine Science, University of Malaga, Málaga, Spain.
The inclusion of microalgae in functional fish diets has a notable impact on the welfare, metabolism and physiology of the organism. The microbial communities associated with the fish are directly influenced by the host's diet, and further understanding the impact on mucosal microbiota is needed. This study aimed to analyze the microbiota associated with the skin and gills of Sparus aurata fed a diet containing 10% microalgae.
View Article and Find Full Text PDFIncreased expression of plasma mir-9-3p and let-7b-3p in methamphetamine use disorder and its clinical significance.
Sci Rep
December 2024
Department of Drug Prohibition and Public Security, Criminal Investigation Police University of China, Shenyang, 110035, China.
Methamphetamine use disorder has emerged as a significant public health concern globally. This study endeavors to elucidate the alterations in expression changes of miRNAs in the plasma of methamphetamine use disorder and elucidate the alterations in miRNA expression in the plasma of individuals with methamphetamine use disorder and investigate the relationship between these differentially expressed miRNAs and the disorder itself, cravings for methamphetamine, and associated mental disorders. Furthermore, the study seeks to clarify the expression of downstream target molecules of specific miRNAs in the plasma of methamphetamine use disorder, assess the diagnostic utility of these miRNAs and their target molecules, explore their potential as biomarkers, and identify potential targets for the diagnosis and treatment of methamphetamine use disorder.
View Article and Find Full Text PDFFast and sensitive multivalent spatial pattern-recognition for circular RNA detection.
Nat Commun
December 2024
Department of Pharmacology, Jiangsu Provincial Key Laboratory of Critical Care Medicine, School of Medicine, Southeast University, Nanjing, China.
While circular RNAs (circRNAs) exhibit lower abundance compared to corresponding linear RNAs, they demonstrate potent biological functions. Nevertheless, challenges arise from the low concentration and distinctive structural features of circRNAs, rendering existing methods operationally intricate and less sensitive. Here, we engineer an intelligent tetrahedral DNA framework (TDF) possessing precise spatial pattern-recognition properties with exceptional sensing speed and sensitivity for circRNAs.
View Article and Find Full Text PDFD-loop mutations in mitochondrial DNA are a risk factor for chemotherapy resistance in esophageal cancer.
Sci Rep
December 2024
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, 2-2-E2, Yamada-Oka, Suita, Osaka, 565-0871, Japan.
Esophageal cancer is a highly aggressive disease, and acquired resistance to chemotherapy remains a significant hurdle in its treatment. mtDNA, crucial for cellular energy production, is prone to mutations at a higher rate than nuclear DNA. These mutations can accumulate and disrupt cellular function; however, mtDNA mutations induced by chemotherapy in esophageal cancer remain unexplored.
View Article and Find Full Text PDFMultivariate stochastic modeling for transcriptional dynamics with cell-specific latent time using SDEvelo.
Nat Commun
December 2024
School of Data Science, The Chinese University of Hong Kong-Shenzhen, Shenzhen, China.
Recently, RNA velocity has driven a paradigmatic change in single-cell RNA sequencing (scRNA-seq) studies, allowing the reconstruction and prediction of directed trajectories in cell differentiation and state transitions. Most existing methods of dynamic modeling use ordinary differential equations (ODE) for individual genes without applying multivariate approaches. However, this modeling strategy inadequately captures the intrinsically stochastic nature of transcriptional dynamics governed by a cell-specific latent time across multiple genes, potentially leading to erroneous results.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!