tRNA-derived fragments (tRFs) are frequently dysregulated in cancers, and approaches for the detection of tRFs within biological samples are vital for their expression analysis and functional exploration. Here, we present a protocol for detecting tRFs using a modified TaqMan quantitative real-time PCR (qRT-PCR)-based technique, Dumbbell-PCR (Db-PCR). We describe steps for primer and adapter design, adapter-RNA ligation, and RNA detection. This protocol streamlines and enhances the precision of tRF quantification in cells, tissues, and plasma, facilitating a time-efficient and reliable assessment of their presence. For complete details on the use and execution of this protocol, please refer to Yu et al. and Sun et al..
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http://dx.doi.org/10.1016/j.xpro.2024.103435 | DOI Listing |
Methods Mol Biol
December 2024
Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger-Center of Molecular Medicine, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
B lymphocytes (B cells) are a type of white blood cell that play an essential role in the adaptive immune response. They are derived from pluripotent hematopoietic stem cells and undergo several developmental stages in the bone marrow and secondary lymphoid organs to become effector cells. B cells can act as antigen-presenting cells, secrete cytokines, generate immunological memory as memory B cells, and produce and secrete high-affinity antibodies as plasma B cells.
View Article and Find Full Text PDFJ Cell Mol Med
December 2024
Department of Respiratory Medicine, The Affiliated Lihuili Hospital of Ningbo University, Zhejiang, China.
Transfer RNA-derived fragments (tRFs) are a new class of small non-coding RNAs. Recent studies suggest that tRFs participate in some pathological processes. However, the biological activities and processes of tRFs in lung cancer cells remain mainly unclear.
View Article and Find Full Text PDFFront Genet
November 2024
Department of Radiation Oncology, Cheeloo College of Medicine, Qilu Hospital of Shandong University, Shandong University, Jinan, Shandong, China.
The main function of transfer RNAs (tRNAs) is to carry amino acids into the ribosome and synthesize proteins under the guidance of messenger RNAs (mRNAs). In addition to this, it has been observed that tRNAs undergo precise cleavage at specific loci, giving rise to an extensive array of distinct small RNAs, termed tRNA-derived small RNAs (tsRNAs). Existing studies have shown that tsRNAs are widely present across various organisms and comprehensively regulate gene expression, aberrant expression of tsRNAs is inextricably linked to tumorigenesis and development, thus, a systematic understanding of tsRNAs is necessary.
View Article and Find Full Text PDFMicroPubl Biol
November 2024
Pharmacology, University of South Alabama College of Medicine, Mobile, AL.
The excision of specific tRNA-derived small RNAs (tsRNAs), yRNA-derived small RNAs (ysRNAs) and ribosomal RNA-derived small RNAs (rsRNAs) is now well established. Several reports have suggested many of these fragments function much like traditional microRNAs (miRNAs). That said, whereas the expressions of the majority of appreciably expressed miRNAs in HCT116 colon cancer cells are significantly decreased in individual knockouts (KOs) of DROSHA, DGCR8, XPO5, and DICER, on average, only 3.
View Article and Find Full Text PDFZhejiang Da Xue Xue Bao Yi Xue Ban
November 2024
School of Medicine, Hangzhou City University, Hangzhou 310015, China.
Neurodegenerative diseases are a heterogeneous group of neurological disorders characterized by the progressive loss of neurons in the central or peripheral nervous system. Research on the pathogenesis and drug targets of these diseases still faces many challenges due to the complex etiology. In recent years, the role of transfer RNA (tRNA) epigenetic modifications in neurodegenerative diseases has attracted widespread attention.
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