Article
Multidisciplinary Sciences
Valeria Genoud, Martin Stortz, Ariel Waisman, Bruno G. Berardino, Paula Verneri, Virginia Dansey, Melina Salvatori, Federico Remes Lenicov, Valeria Levi
Summary: The PK+HID method is an extraction-free RT-qPCR technique for SARS-CoV-2 detection, showing improved performance compared to traditional heat-inactivation procedures. It can be combined with multiplexing RT-qPCR kits, demonstrating high sensitivity and specificity in clinical sample analysis.
Article
Multidisciplinary Sciences
Amy Oreskovic, Barry R. Lutz
Summary: The study developed an ultrasensitive urine cfDNA extraction method that efficiently recovers short cfDNA fragments, achieving high sensitivity and detection limits. By combining hybridization with ultra-short qPCR design, it is possible to efficiently capture and amplify cfDNA fragments as short as 25 bp.
Article
Engineering, Chemical
Rita Carapito, Sandra C. Bernardo, Matheus M. Pereira, Marcia C. Neves, Mara G. Freire, Fani Sousa
Summary: Nucleic acids, especially RNA, have been explored as potential biopharmaceuticals for gene therapy. However, their labile nature poses challenges in their production and purification. This study investigates the use of chromatographic silica-based supports functionalized with ionic liquids for RNA purification. The results show promising potential for the development of a more efficient and safer method for RNA isolation.
SEPARATION AND PURIFICATION TECHNOLOGY
(2023)
Article
Chemistry, Analytical
Saki Matsumoto, Shuntaro Takahashi, Sudipta Bhowmik, Tatsuya Ohyama, Naoki Sugimoto
Summary: Hydration around nucleic acids is crucial for their stability and interaction with binding molecules. The study demonstrates that the core G4 helix region in G-quadruplex (G4) RNA formation is more dehydrated compared to other parts. These findings help in predicting the binding of G4 ligands and provide information for molecular interactions during G4 formation in crowded cellular environments.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Ya-Ling Tan, A-Qian Huang, Li-Juan Tang, Jian-Hui Jiang
Summary: The development of multiplexed droplet loop-mediated isothermal amplification (multiplexed dLAMP) technique, which allows simultaneous quantification of multiple targets, provides a highly sensitive digital nucleic acid method for rapid and effective detection of viruses and other pathogens. The technology not only has been successfully demonstrated in the laboratory, but also holds promise for digital detection of HCV and HIV RNA in clinical applications. With high precision and low detection limits, it offers a new approach for the prevention and control of infectious diseases.
Review
Engineering, Biomedical
Ping Song, Lucia Ruojia Wu, Yan Helen Yan, Jinny X. Zhang, Tianqing Chu, Lawrence N. Kwong, Abhijit A. Patel, David Yu Zhang
Summary: This Review discusses the challenges and opportunities of technologies for the analysis of cell-free DNA for cancer diagnosis. It highlights the potential of cfDNA as biomarkers for guiding therapy, monitoring drug resistance, and early detection of cancers. However, the low concentrations and fragmented nature of cfDNA present challenges for clinical diagnostic applications. The Review describes state-of-the-art technologies and highlights the need to address biological and technical challenges to improve cancer diagnostics and patient care.
NATURE BIOMEDICAL ENGINEERING
(2022)
Article
Nanoscience & Nanotechnology
Bradley M. Downs, Saraswati Sukumar
Summary: Many studies have shown that blood-based liquid biopsies can detect cancer in its early stages, but the limitation lies in the volume of blood required. Researchers have developed a new technology that can capture ctDNA from flowing unaltered plasma, but further validation and optimization are needed.
ACS APPLIED MATERIALS & INTERFACES
(2022)
Article
Biochemistry & Molecular Biology
Julian M. Hess, William K. Jannen, Daniel P. Aalberts
Summary: This study introduces a model to describe the unfolding costs of the binding site and implements it in an O(N) algorithm called BINDOLIGONET. The research reveals significant differences in the free energy parameters for the four bases. The accuracy of donor splice site prediction improves when using their calculation method, and the base-dependent free energies are also correlated with efficient ribosome docking near the start codon.
JOURNAL OF MOLECULAR BIOLOGY
(2022)
Article
Chemistry, Analytical
Ya-Ling Tan, Hongjian Chen, Zhen-Kun Wu, Jianjun He, Jian-Hui Jiang
Summary: The study developed a sensitive HEADLAMP method based on HpaII for accurate detection of DNA methylation levels. Compared to traditional methods, HEADLAMP can more accurately detect low levels of DAPK1 methylation and show potential clinical applications in clinical tissue samples.
ANALYTICAL CHEMISTRY
(2021)
Review
Biochemical Research Methods
Mojca Milavec, Megan H. Cleveland, Young-Kyung Bae, Robert Wielgosz, Maxim Vonsky, Jim F. Huggett
Summary: Nucleic acid analysis is widely used in various fields such as medicine, food safety, and environmental monitoring, but users are often unaware of the global metrological infrastructure. International efforts, particularly through the NAWG, focus on improving measurement capabilities and service dissemination to enhance the reliability, comparability, and traceability of nucleic acid measurement results.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)
Article
Biochemical Research Methods
Steven Chin, Alexandre Goyon, Kelly Zhang, Kenji L. Kurita
Summary: Accurate sequencing of sgRNAs is crucial for CRISPR/Cas9 genome editing, and a method using protective DNA primers and RNase T1 digestion coupled with HILIC-HRMS was developed. Without RNase digestion, top-down mass spectrometry struggles to fragment precursor ions in large RNA oligonucleotides. However, combining primer protection with RNase T1 digestion allows for full sgRNA sequence coverage by providing large enough digestion products to prevent redundancy and small enough products for base resolution by tandem mass spectrometry. This technique has the potential to expedite accurate sequence confirmation of chemically modified sgRNA oligonucleotides.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2023)
Review
Biotechnology & Applied Microbiology
Xiaowei Ma, Jingsong Xu, Fei Zhou, Jing Ye, Donglei Yang, Hua Wang, Pengfei Wang, Min Li
Summary: This review highlights recent advances in PCR-free nucleic acid detection methods for SARS-CoV-2, including emerging methods of isothermal amplification, nucleic acid enzymes, electrochemistry, and CRISPR.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Jun Gao, Zhaofeng Gao, Andrea A. Putnam, Alicia K. Byrd, Sarah L. Venus, John C. Marecki, Andrea D. Edwards, Haley M. Lowe, Eckhard Jankowsky, Kevin D. Raney
Summary: The interaction between G-quadruplex DNA and the DEAD-box helicase Ded1p's N-terminal intrinsically disordered domain has been shown to decrease RNA unwinding activity while enhancing liquid-liquid phase separation, both in vitro and in cells. This study highlights the complex effects of quadruplex DNA on an enzyme with intrinsically disordered domains.
CHEMICAL COMMUNICATIONS
(2021)
Review
Biophysics
Alberto Marin-Gonzalez, J. G. Vilhena, Ruben Perez, Fernando Moreno-Herrero
Summary: Understanding the complex mechanical behavior of DNA at different scales requires taking a molecular perspective, which recent single-molecule experiments and molecular dynamics simulations have provided insights into. These studies have identified sequence-dependent properties, effects of cytosine methylation and DNA mismatches, as well as differences in mechanical properties between DNA and double-stranded RNA. Thorough examination of single-molecule literature has led to the establishment of general 'rules' explaining the mechanics of nucleic acids at the base pair level, which can be valuable for designing DNA and RNA nanostructures in the future.
QUARTERLY REVIEWS OF BIOPHYSICS
(2021)
Article
Chemistry, Analytical
Miranda N. Emaus, Jared L. Anderson
Summary: The study investigated three hydrophobic magnetic ionic liquids (MILs) as EXPAR additives and miRNA extraction solvents. The addition of MIL to the EXPAR buffer decreased non-specific amplification background signal and increased reaction rate. Reactions containing MIL could detect miRNA at concentration levels down to 10 aM.
ANALYTICA CHIMICA ACTA
(2021)
Review
Chemistry, Analytical
Kevin D. Clark, Miranda N. Emaus, Marcelino Varona, Ashley N. Bowers, Jared L. Anderson
JOURNAL OF SEPARATION SCIENCE
(2018)
Article
Biochemical Research Methods
Kevin D. Clark, Maria J. Trujillo-Rodriguez, Jared L. Anderson
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2018)
Editorial Material
Biochemical Research Methods
Jared L. Anderson, Kevin D. Clark
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2018)
Article
Chemistry, Analytical
Xiong Ding, Kevin D. Clark, Marcelino Varona, Miranda N. Emaus, Jared L. Anderson
ANALYTICA CHIMICA ACTA
(2019)
Article
Chemistry, Analytical
Stephanie A. Hice, Kevin D. Clark, Jared L. Anderson, Byron F. Brehm-Stecher
ANALYTICAL CHEMISTRY
(2019)
Article
Chemistry, Analytical
Kevin D. Clark, Chenghui Zhu, Jared L. Anderson
ANALYTICAL CHEMISTRY
(2019)
Article
Chemistry, Analytical
Kevin D. Clark, Marina C. Philip, Yanqi Tan, Jonathan Sweedler
ANALYTICAL CHEMISTRY
(2020)
Article
Chemistry, Multidisciplinary
Kevin D. Clark, Colin Lee, Rhanor Gillette, Jonathan Sweedler
Summary: The study suggests that subtle changes in post-transcriptional modifications may regulate central nervous system plasticity and activity-induced protein synthesis. Using mass spectrometry, researchers found unique RNA modification patterns in neurons during learning processes, with two tRNA modifications showing significantly higher levels in trained animals. This indicates a link between noncoding RNA modifications and non-associative learning.
ACS CENTRAL SCIENCE
(2021)
Article
Chemistry, Analytical
Kevin D. Clark, Stanislav S. Rubakhin, Jonathan Sweedler
Summary: SNRMA-MS is the first analytical approach capable of simultaneously quantifying numerous RNA modifications in single neurons and revealing cell-specific modification profiles.
ANALYTICAL CHEMISTRY
(2021)
Article
Genetics & Heredity
Madeline E. Melzer, Jonathan Sweedler, Kevin D. Clark
Summary: The reversible and substoichiometric modification of RNA has been recognized as an additional layer of translational regulation. A nuclease protection assay coupled with MALDI-MS was developed to rapidly characterize the modification stoichiometry of consensus motifs, providing a promising approach for studying RNA modifications.
Article
Multidisciplinary Sciences
Kevin D. Clark, Stanislav S. Rubakhin, Jonathan Sweedler
Summary: Post-transcriptional modifications (PTMs) of RNA play a role in regulating translation in the central nervous system (CNS). A protocol called single-neuron RNA modification analysis by mass spectrometry (SNRMA-MS) is described, which can detect and quantify multiple RNA modifications in individual neurons. This method has been validated in A. californica, and has revealed the diverse RNA modification patterns in identified neurons.
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
(2022)
Article
Biochemical Research Methods
Arya Patel, Kevin D. Clark
Summary: Post-transcriptional modifications to RNA play a crucial role in regulating translation in the CNS. Analytical methods such as RNA sequencing and LC-MS/MS have provided insights into the landscape of RNA modifications in bulk CNS tissues. However, recent advancements in techniques for single-cell and subcellular detection have revealed heterogeneity in RNA modification profiles, raising questions about cell-specific distributions and functions of modified nucleosides in the brain.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2023)
Article
Physics, Atomic, Molecular & Chemical
Max D. Sharin, Gabriella M. Floro, Kevin D. Clark
Summary: Electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) have revolutionized nucleic acid analysis by mass spectrometry. However, complex biological samples and large nucleic acids pose challenges for these techniques. Over the past two decades, innovations in sample preparation methods have been vital in overcoming these challenges and advancing high-throughput, sensitive, and broadly applicable MS methods for nucleic acid analysis.
INTERNATIONAL JOURNAL OF MASS SPECTROMETRY
(2023)
Article
Chemistry, Analytical
Kevin D. Clark
Summary: This article discusses the challenges and opportunities in the field of RNA modifications, with a focus on advancements in sample preparation and microextraction methods. These techniques enable the analysis of RNA modifications in single cells, providing new insights into their function in the brain and biological systems.
LC GC NORTH AMERICA
(2022)