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Synthetic Lesions with a Fluorescein Carbamoyl Group As Analogs of Bulky Lesions Removable by Nucleotide Excision Repair: A Comparative Study on Properties.
Acta Naturae
January 2024
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Sciences, Novosibirsk, 630090 Russian Federation.
Mammalian nucleotide excision repair (NER), known for its broad substrate specificity, is responsible for removal of bulky lesions from DNA. Over 30 proteins are involved in NER, which includes two distinct pathways: global genome NER and transcription-coupled repair. The complexity of these processes, the use of extended DNA substrates, and the presence of bulky DNA lesions induced by chemotherapy have driven researchers to seek more effective methods by which to assess NER activity, as well as to develop model DNAs that serve as efficient substrates for studying lesion removal.
View Article and Find Full Text PDFFluorescent primers amplification refractory mutation system qPCR (FP ARMS-qPCR) for MTHFR C677T SNP genotyping.
Mol Biol Rep
November 2024
Second School of Clinical Medicine, Henan University of Chinese Medicine, Zhengzhou, 450002, China.
Background: The currently used methods for the detection of methylene tetrahydrofolic acid reductase (MTHFR) C677T single nucleotide polymorphism (SNP) are either time-consuming or expensive. In this study, we devised an accurate, rapid and easy-to-use SNP detection system based on fluorescent primers amplification refractory mutation system qPCR, known as FP ARMS-qPCR.
Methods: Fluorescent primers (FPs) modified by fluorescent dye or quencher near the 3' terminal thymine were designed.
Characterization of Hibiscus Chlorotic Ringspot Virus-Derived vsiRNAs from Infected Hibiscus rosa-sinensis in China.
Plant Pathol J
October 2024
Key Laboratory of Landscape Plants with Fujian and Taiwan Characteristics of Fujian Colleges and Universities, School of Biological Sciences and Biotechnology, Minnan Normal University, Zhangzhou 363000, China.
Article Synopsis
- Progress has been made in understanding the relationship between Hibiscus rosa-sinensis and hibiscus chlorotic ringspot virus (HCRSV), but many interactions are still not well understood.
- HCRSV infection in H. rosa-sinensis was confirmed using various advanced techniques, and the virus was found to have a 3,909 nucleotide long genome with features similar to other closely related viruses.
- The study identified specific viral small interfering RNAs (vsiRNAs) derived from HCRSV, discovering that they predominantly come from the positive-strand RNA and exhibit unique patterns in distribution which may relate to the secondary structures of the viral genome.
Templated 3' terminal fluorescent labeling of RNA using Klenow DNA polymerase.
MethodsX
December 2024
Saint Joseph's University, Philadelphia, PA, USA.
A long-standing challenge in the study of RNA structure-function dynamics using fluorescence-based methods has been the precise attachment of fluorophores to structured RNA molecules. Despite significant advancements in the field, existing techniques have limitations, especially for 3' end labeling of long, structured RNAs. In response to this challenge, we developed a chemo-enzymatic method that uses Klenow DNA polymerase to label RNAs.
View Article and Find Full Text PDFPrecise Insertion of AttB Sequences in Goat Genome Using Enhanced Prime Editor.
Int J Mol Sci
August 2024
Key Laboratory of Livestock Biology, Northwest A&F University, Yangling 712100, China.
Prime editor, an editing tool based on the CRISPR/Cas9 system, allows for all 12 types of nucleotide exchanges and arbitrary indels in genomic sequences without the need for inducing DNA double-strand breaks. Despite its flexibility and precision, prime editing efficiency is still low and hindered by various factors such as target sites, editing types, and the length of the primer binding site. In this study, we developed a prime editing system by incorporating an RNA motif at the 3' terminal of the pegRNA and integrating all twin prime editor factors into a single plasmid.
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