scholarly journals Acidification of the Oxygen Scavenging System in Single-Molecule Fluorescence Studies: In Situ Sensing with a Ratiometric Dual-Emission Probe

2010 ◽  
Vol 82 (14) ◽  
pp. 6132-6138 ◽  
Author(s):  
Xinghua Shi ◽  
John Lim ◽  
Taekjip Ha
Keyword(s):  
Single Molecule ◽  
Single Molecule Fluorescence ◽  
Dual Emission ◽  
Fluorescence Studies ◽  
Oxygen Scavenging ◽  
In Situ Sensing

scholarly journals Single-molecule fluorescence studies of RNA: A decade’s progress

2009 ◽  
Vol 476 (1-3) ◽  
pp. 1-10 ◽  
Author(s):  
Krishanthi S. Karunatilaka ◽  
David Rueda
Keyword(s):  
Single Molecule ◽  
Single Molecule Fluorescence ◽  
Fluorescence Studies

scholarly journals In situ genotyping of a pooled strain library after characterizing complex phenotypes

2017 ◽  
Author(s):  
Michael J. Lawson ◽  
Daniel Camsund ◽  
Jimmy Larsson ◽  
Özden Baltekin ◽  
David Fange ◽  
...  
Keyword(s):  
Fluorescence Imaging ◽  
Single Molecule ◽  
Single Molecule Fluorescence ◽  
Detailed Characterization ◽  
E Coli ◽  
Complex Phenotypes

So far, it has not been possible to perform advanced microscopy on pool generated strain libraries and at the same time know each strain’s genotype. We have overcome this barrier by identifying the genotypes for individual cells in situ after a detailed characterization of the phenotype. The principle is demonstrated by single molecule fluorescence imaging of E. coli strains harboring barcoded plasmids that express a sgRNA which suppress different genes through dCas9.

scholarly journals Single-molecule fluorescence studies on cotranscriptional G-quadruplex formation coupled with R-loop formation

2020 ◽  
Vol 48 (16) ◽  
pp. 9195-9203
Author(s):  
Gunhyoung Lim ◽  
Sungchul Hohng
Keyword(s):  
Gene Regulation ◽  
Single Molecule ◽  
Genome Instability ◽  
Feedback Mechanism ◽  
Rnase H ◽  
Formation Mechanisms ◽  
Loop Formation ◽  
Single Molecule Fluorescence ◽  
Fluorescence Studies ◽  
G Quadruplex

Abstract G-quadruplex (GQ) is formed at various regions of DNA, including telomeres of chromosomes and regulatory regions of oncogenes. Since GQ is important in both gene regulation and genome instability, the biological and medical implications of this abnormal DNA structure have been intensively studied. Its formation mechanisms, however, are not clearly understood yet. We report single-molecule fluorescence experiments to monitor the cotranscriptional GQ formation coupled with R-loop formation using T7 RNA polymerase. The GQ is formed very rarely per single-round transcription. R-loop formation precedes and facilitates GQ formation. Once formed, some GQs are extremely stable, resistant even to RNase H treatment, and accumulate in multiple-round transcription conditions. On the other hand, GQ existing in the non-template strand promotes the R-loop formation in the next rounds of transcription. Our study clearly shows the existence of a positive feedback mechanism of GQ and R-loop formations, which may possibly contribute to gene regulation and genome instability.

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