ArticleNucleic acids research2018
Capping-RACE: a simple, accurate, and sensitive 5' RACE method for use in prokaryotes.
Article in Nucleic acids research, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 22 citations in OpenAlex.
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- Pervasive transcription enhances the accessibility of H-NS-silenced promoters and generates bistability inProceedings of the National Academy of Sciences of the United States of America · 2022Article
- Exploring clinical implications and role of non-coding RNAs in lung carcinogenesis.Molecular biology reports · 2022Review
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- Role of the Two-Component System CiaRH in the Regulation of Efflux Pump SatAB and Its Correlation with Fluoroquinolone Susceptibility.Microbiology spectrum · 2022Article
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- An Update of Long-Noncoding RNAs in Acute Kidney Injury.Frontiers in physiology · 2022Review
- The Connection between Czc and Cad Systems Involved in Cadmium Resistance inInternational journal of molecular sciences · 2021Article
- Determination of the vRNA and cRNA promoter activity by M segment-specific non-coding nucleotides of influenza A virus.RNA biology · 2021Article
- Global Regulator of Rubber Degradation in Gordonia polyisoprenivorans VH2: Identification and Involvement in the Regulation Network.Applied and environmental microbiology · 2020Article
- AP-TSS: A New Method for the Analysis of RNA Expression from Particular and Challenging Transcription Start Sites.Biomolecules · 2020Article
- Long Non-Coding RNAs: The Regulatory Mechanisms, Research Strategies, and Future Directions in Cancers.Frontiers in oncology · 2020Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Rapid amplification of cDNA ends (RACE) is a prevalent technique used to obtain the 5' ends of transcripts. Several different 5' RACE methods have been developed, and one particularly simple and efficient approach called CapFinder relies on the 5' cap-dependent template-switching that occurs in eukaryotes. However, most prokaryotic transcripts lack a 5' cap structure. Here, we report a procedure to capture primary transcripts based on capping the 5' triphosphorylated RNA in prokaryotes. Primary transcripts were first treated with vaccinia capping enzyme to add a 5' cap structure. First-strand cDNA was then synthesized using Moloney murine leukaemia virus reverse transcriptase. Finally, a template-switching oligonucleotide with a tail containing three ribonucleic acid guanines was hybridized to the cDNA 3' poly(C) and further used as template for reverse transcriptase. It is oligonucleotide sequence independent and is more sensitive compared to RLM-RACE. This approach specifically identified the transcription start sites of ompA, sodB and shiA in Escherichia coli and of ompA, rne and rppH in Brucella melitensis. Furthermore, we also successfully identified the transcription start sites of small noncoding genes ryhB and micC in E. coli and bsnc135 and bsnc149 in B. melitensis. Our findings suggest that Capping-RACE is a simple, accurate, and sensitive 5' RACE method for use in prokaryotes.
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