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lncRNA - Long Non-coding RNAs02:39

lncRNA - Long Non-coding RNAs

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In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...
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Epigenetics is the study of inherited changes in a cell's phenotype without changing the DNA sequences. It provides a form of memory for the differential gene expression pattern to maintain cell lineage, position-effect variegation, dosage compensation, and maintenance of chromatin structures such as telomeres and centromeres. For example, the structure and location of the centromere on chromosomes are epigenetically inherited. Its functionality is not dictated or ensured by the underlying...
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The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
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Visualizing Long Noncoding RNAs on Chromatin.

Michael Hinten1, Emily Maclary1, Srimonta Gayen1

  • 1Department of Human Genetics, University of Michigan Medical School, 3710-D Med. Sci II SPC 5618, 1241 E. Catherine Street, Ann Arbor, MI, 48109, USA.

Methods in Molecular Biology (Clifton, N.J.)
|January 2, 2016
PubMed
Summary
This summary is machine-generated.

This study presents advanced fluorescence in situ hybridization (FISH) protocols for highly sensitive detection of RNA, DNA, and proteins in single cells. These methods link long noncoding RNA (lncRNA) function to epigenetic effects.

Keywords:
ChromatinDNA FISHEpigeneticFluorescence in situ hybridizationHistone modificationsImmunofluorescenceLong noncoding RNAsRNA FISH

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Fluorescence in situ hybridization (FISH) is a powerful technique for detecting nucleic acid sequences in cells.
  • RNA FISH combined with protein detection offers insights into long noncoding RNA (lncRNA) function and epigenetic effects.
  • Previous limitations in detecting low-abundance or unstable lncRNAs are being overcome by advanced in situ techniques.

Purpose of the Study:

  • To describe an integrated set of protocols for sensitive detection of specific RNAs, DNAs, proteins, and histone modifications in single cells.
  • To enable the investigation of lncRNA function by connecting them to epigenetic modifications.
  • To facilitate the study of lncRNAs expressed at low levels or that are unstable.

Main Methods:

  • Development of highly sensitive FISH-based protocols for simultaneous detection of RNA, DNA, proteins, and histone modifications.
  • Utilizing conventional fluorescence microscopy for visualization.
  • Integrated approach allowing individual or combined detection of multiple targets within single cells.

Main Results:

  • Demonstration of a high level of sensitivity in detecting various molecular targets within single cells.
  • Establishment of protocols capable of visualizing low-abundance or unstable lncRNAs.
  • Facilitation of the connection between lncRNAs and epigenetic effects through combined detection methods.

Conclusions:

  • The described integrated protocols significantly advance the ability to study lncRNA function and epigenetic roles at the single-cell level.
  • These methods provide a sensitive and versatile tool for molecular and cell biology research.
  • The approach opens new avenues for understanding gene regulation and epigenetic mechanisms involving lncRNAs.