Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

6.6K
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
6.6K
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

3.5K
3.5K
Cell Specific Gene Expression01:58

Cell Specific Gene Expression

16.5K
Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
16.5K
pre-mRNA Processing02:01

pre-mRNA Processing

57.4K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
57.4K
Regulated mRNA Transport02:22

Regulated mRNA Transport

7.0K
In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
7.0K
Nuclear Export of mRNA02:31

Nuclear Export of mRNA

8.8K
Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
8.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Combining multiplexed functional data to improve variant classification.

Genome medicine·2026
Same author

Long-Term Outcomes in Patients With Recurrent Ovarian Cancer and Exceptional Response to PARP Inhibitors.

JAMA oncology·2026
Same author

Homologous recombination deficiency-driven genomic instability in ovarian cancer as an indicator of BRCA1 and BRCA2 variant pathogenicity.

American journal of human genetics·2026
Same author

Updated ENIGMA recommendations for reporting germline variants in cancer susceptibility genes and their translation into twenty languages.

Journal of medical genetics·2026
Same author

Bayesian Analysis of Postoperative Complication Risk Associated With Preoperative Exposure to Fine Particulate Matter: A Single-Center Cohort Study.

Acta anaesthesiologica Scandinavica·2026
Same author

Investigating the contribution of rare non-coding variants in BRCA1, BRCA2 and PALB2 to hereditary breast cancer.

NPJ breast cancer·2026

Related Experiment Video

Updated: Jan 29, 2026

Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1
08:53

Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1

Published on: February 17, 2011

15.1K

Quantifying BRCA1 and BRCA2 mRNA Isoform Expression Levels in Single Cells.

Vanessa L Lattimore1, John F Pearson2, Arthur E Morley-Bunker3

  • 1Department of Pathology and Biomedical Science, University of Otago, Christchurch 8011, New Zealand. vanessa.lattimore@otago.ac.nz.

International Journal of Molecular Sciences
|February 10, 2019
PubMed
Summary

This study introduces a single-cell RNA in situ hybridization method to analyze BRCA1/2 splicing variants. It reveals stochastic mRNA expression, impacting previous cancer risk assessments for these variants.

Keywords:
BRCA1BRCA2RNAscopemRNA expressionsingle cellsplicing

More Related Videos

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
09:22

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Published on: February 28, 2021

6.0K
Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence
06:25

Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence

Published on: February 10, 2023

2.5K

Related Experiment Videos

Last Updated: Jan 29, 2026

Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1
08:53

Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1

Published on: February 17, 2011

15.1K
Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors
09:22

Functional Assessment of BRCA1 variants using CRISPR-Mediated Base Editors

Published on: February 28, 2021

6.0K
Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence
06:25

Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence

Published on: February 10, 2023

2.5K

Area of Science:

  • Genetics
  • Molecular Biology
  • Cancer Research

Background:

  • BRCA1 and BRCA2 spliceogenic variants are linked to increased breast and ovarian cancer risks.
  • Traditional analyses of splicing patterns lack single-cell resolution, potentially obscuring cellular variation.
  • Understanding variant impact on mRNA expression is crucial for accurate cancer risk assessment.

Purpose of the Study:

  • To develop and validate a single-cell RNA in situ hybridization method for analyzing BRCA1 and BRCA2 mRNA splicing events.
  • To investigate the expression patterns of specific spliceogenic variants (BRCA1c.671-2 A>G and BRCA2c.7988 A>T) at the single-cell level.
  • To compare single-cell findings with traditional population-based analyses.

Main Methods:

  • Utilized RNA in situ hybridization to quantify absolute mRNA splicing events in single lymphoblastoid cells.
  • Analyzed cells with known spliceogenic variants (BRCA1c.671-2 A>G, BRCA2c.7988 A>T) and control cells.
  • Measured expression levels and proportions of targeted BRCA1 and BRCA2 mRNA isoforms.

Main Results:

  • A significant proportion of cells (>42%) showed no detectable mRNA expression for the targeted genes.
  • Increased levels of BRCA2 ∆17_18 mRNA were observed in cells with the BRCA2c.7988 A>T variant.
  • Cells with BRCA1c.671-2 A>G did not show significantly increased BRCA1 ∆11 levels, but a higher proportion expressed the targeted splicing event compared to controls.

Conclusions:

  • BRCA1/2 mRNA expression is stochastic at the single-cell level, challenging previous RT-PCR findings.
  • Single-cell analysis provides a more nuanced understanding of spliceogenic variant effects on mRNA isoforms.
  • Further research is needed to establish this method for diagnostic applications in spliceogenic variant assessment.