Chemo-biological mRNA imaging with single nucleotide specificity

2019 ◽  
Vol 55 (98) ◽  
pp. 14817-14820 ◽  
Author(s):  
Andrea Knoll ◽  
Svenja Kankowski ◽  
Sophie Schöllkopf ◽  
Jochen C. Meier ◽  
Oliver Seitz
Keyword(s):  
Cellular Imaging ◽  
Rna Detection ◽  
Single Nucleotide ◽  
Imaging Methods ◽  
Combined Use ◽  
Rna Imaging ◽  
Nucleotide Specificity

The combined use of “biological” RNA imaging methods based on MS2 technology and “chemical” RNA detection by FIT probes allows unambiguous cellular imaging of a C → U edit in mRNA encoding for GlyR α2.

scholarly journals Advanced Circuit and Cellular Imaging Methods in Nonhuman Primates

2019 ◽  
Vol 39 (42) ◽  
pp. 8267-8274
Author(s):  
Stephen L. Macknik ◽  
Robert G. Alexander ◽  
Olivya Caballero ◽  
Jordi Chanovas ◽  
Kristina J. Nielsen ◽  
...  
Keyword(s):  
Nonhuman Primates ◽  
Cellular Imaging ◽  
Imaging Methods

scholarly journals Cellular microRNA detection with miRacles: microRNA- activated conditional looping of engineered switches

2019 ◽  
Vol 5 (3) ◽  
pp. eaau9443 ◽  
Author(s):  
Arun Richard Chandrasekaran ◽  
Molly MacIsaac ◽  
Paromita Dey ◽  
Oksana Levchenko ◽  
Lifeng Zhou ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Northern Blotting ◽  
Single Step ◽  
Single Nucleotide ◽  
Disease Biomarkers ◽  
Microrna Detection ◽  
Detection Assay ◽  
Polymerase Chain ◽  
Regulatory Rnas ◽  
Nucleotide Specificity

MicroRNAs are short noncoding regulatory RNAs that are increasingly used as disease biomarkers. Detection of microRNAs can be arduous and expensive and often requires amplification, labeling, or radioactive probes. Here, we report a single-step, nonenzymatic microRNA detection assay using conformationally responsive DNA nanoswitches. Termed miRacles (microRNA-activated conditional looping of engineered switches), our assay has subattomole sensitivity and single-nucleotide specificity using an agarose gel electrophoresis readout. We detect cellular microRNAs from nanogram-scale RNA extracts of differentiating muscle cells and multiplex our detection for several microRNAs from one biological sample. We demonstrate 1-hour detection without expensive equipment or reagents, making this assay a compelling alternative to quantitative polymerase chain reaction and Northern blotting.

scholarly journals Deep-red fluorogenic cyanine dyes carrying an amino group-terminated side chain for improved RNA detection and nucleolar RNA imaging

RSC Advances ◽  
2021 ◽  
Vol 11 (56) ◽  
pp. 35436-35439
Author(s):  
Yusuke Sato ◽  
Yugo Igarashi ◽  
Michiyuki Suzuki ◽  
Kei Higuchi ◽  
Seiichi Nishizawa
Keyword(s):  
Cyanine Dyes ◽  
Cyanine Dye ◽  
Side Chain ◽  
Amino Group ◽  
Rna Detection ◽  
Rna Imaging ◽  
Nucleolar Rna ◽  
Nucleolar Rnas ◽  
In Cells

The introduction of an amino-group-terminated side chain into deep-red emissive benzo[c,d]indole–quinoline monomethine cyanine dye has led to the improved detection of RNAs as well as the imaging of nucleolar RNAs in cells.

Achieving Single-Nucleotide Specificity in Direct Quantitative Analysis of Multiple MicroRNAs (DQAMmiR)

2016 ◽  
Vol 88 (4) ◽  
pp. 2472-2477 ◽  
Author(s):  
David W. Wegman ◽  
Farhad Ghasemi ◽  
Alexander S. Stasheuski ◽  
Anna Khorshidi ◽  
Burton B. Yang ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Single Nucleotide ◽  
Nucleotide Specificity

A novel microfluidic RNA chip for direct, single-nucleotide specific, rapid and partially-degraded RNA detection

Talanta ◽  
2021 ◽  
pp. 122974
Author(s):  
Shun Zhang ◽  
Jiuyi Chen ◽  
Dan Liu ◽  
Bei Hu ◽  
Guangcheng Luo ◽  
...  
Keyword(s):  
Rna Detection ◽  
Single Nucleotide

scholarly journals The Roles of MicroRNAs in Male Infertility

2021 ◽  
Vol 22 (6) ◽  
pp. 2910
Author(s):  
Madalina Gabriela Barbu ◽  
Dana Claudia Thompson ◽  
Nicolae Suciu ◽  
Silviu Cristian Voinea ◽  
Dragos Cretoiu ◽  
...  
Keyword(s):  
Male Infertility ◽  
Recent Literature ◽  
Detection Methods ◽  
Nucleotide Polymorphism ◽  
Contraceptive Methods ◽  
Aberrant Expression ◽  
Rna Detection ◽  
Single Nucleotide ◽  
Future Studies ◽  
Potential Cancer

MicroRNAs applications were vastly studied throughout the years, spanning from potential cancer biomarkers to targeted therapies for various diseases. Out of these utilizations, this paper focuses on their role in male infertility. Approximately 10–15% of worldwide couples are affected by infertility. Out of these, 50% are due to male determinants. The majority of cases still have an undetermined cause. Previous studies have found that the aberrant expression of microRNAs could be linked to certain reproductive dysfunctions in males. Further on, this study looked into the most recent literature published on this subject in order to assess the connection between the up-/down-regulation of various microRNAs and the roles they play in male infertility. MicroRNAs were found to be abundant and stable in the seminal liquid, which led to a facile identification using regular RNA detection methods. It was observed that the concentration of microRNAs in semen was modified in the case of patients suffering from asthenozoospermia and azoospermia. Moreover, idiopathic male infertility was associated with a single nucleotide polymorphism of the microRNA binding site. Future studies should focus their attention on discovering future treatments against male infertility targeting specific microRNAs and also on developing new and improved contraceptive methods.

scholarly journals Influence of the FCGR2A rs1801274 and FCGR3A rs396991 Polymorphisms on Response to Abatacept in Patients with Rheumatoid Arthritis

2021 ◽  
Vol 11 (6) ◽  
pp. 573
Author(s):  
Noelia Márquez Pete ◽  
María del Mar Maldonado Montoro ◽  
Cristina Pérez Ramírez ◽  
Fernando Martínez Martínez ◽  
Juan Enrique Martínez de la Plata ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Concomitant Disease ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Lower Baseline ◽  
Combined Use ◽  
Genes Encoding ◽  
Observational Cohort ◽  
Caucasian Patients ◽  
Encoding Protein

Abatacept (ABA) is an immunosuppressant indicated for treatment of rheumatoid arthritis (RA). Effectiveness might be influenced by clinical RA variants and single-nucleotide polymorphisms (SNPs) in genes encoding protein FCGR2A (His131Arg) and FCGR3A (Phe158Val) involved in pharmacokinetics of ABA. An observational cohort study was conducted in 120 RA Caucasian patients treated with ABA for 6 and 12 months. Patients with the FCGR2A rs1801274-AA genotype (FCGR2A-p.131His) showed a better EULAR response (OR = 2.43; 95% CI = 1.01–5.92) at 12 months and low disease activity (LDA) at 6 months (OR = 3.16; 95% CI = 1.19–8.66) and 12 months (OR = 6.62; 95% CI = 1.25–46.89) of treatment with ABA. A tendency was observed towards an association between the FCGR3A rs396991-A allele (FCGR3A-p.158Phe) and better therapeutic response to ABA after 12 months of treatment (p = 0.078). Moreover, we found a significant association between the low-affinity FCGR2A/FCGR3A haplotypes variable and LDA after 12 months of ABA treatment (OR = 1.59; 95% CI = 1.01–2.58). The clinical variables associated with better response to ABA were lower age at starting ABA (OR = 1.06; 95% CI = 1.02–1.11) and greater duration of ABA treatment (OR = 1.02; 95% CI = 1.01–1.04), lower duration of previous biological therapies (OR = 0.99; 95% CI = 0.98–0.99), non-administration of concomitant disease-modifying antirheumatic drugs (DMARDs) (OR = 24.53; 95% CI = 3.46–523.80), non-use of concomitant glucocorticoids (OR = 0.12; 95% CI = 0.02–0.47), monotherapy (OR = 19.22; 95% CI = 2.05–343.00), lower initial patient’s visual analogue scale (PVAS) value (OR = 0.95; 95% CI = 0.92–0.97), and lower baseline ESR (OR = 0.92; 95% CI = 0.87–0.97). This study showed that high-affinity FCGR2A-p.131His variant, low-affinity FCGR3A-p.158Phe variant, and combined use of FCGR2A/FCGR3A genetic variations could affect ABA effectiveness. Further studies will be required to confirm these results.

scholarly journals Droplet-digital Cas13a assay enables direct single-molecule microRNA quantification

2019 ◽  
Author(s):  
Tian Tian ◽  
Bowen Shu ◽  
Lei Liu ◽  
Xiaoming Zhou
Keyword(s):  
Cell Lines ◽  
Single Molecule ◽  
Absolute Quantification ◽  
Single Nucleotide ◽  
Single Molecule Level ◽  
Broad Applicability ◽  
Precise Quantification ◽  
Nucleotide Specificity ◽  
Direct Quantification

The direct quantification of microRNA at the single-molecule level is still challenging. Herein, we developed a droplet-digital Cas13a assay (ddCA) as a general approach for the amplification-free and absolute quantification of single unlabeled miRNA molecules with single-nucleotide specificity. We demonstrate its simplicity, precise quantification capability, excellent specificity and broad applicability by analyzing microRNAs from synthetic and cell lines-derived materials.

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