Related Experiment Video

Updated: Mar 1, 2026

Analysis of mRNA Nuclear Export Kinetics in Mammalian Cells by Microinjection
11:32

Analysis of mRNA Nuclear Export Kinetics in Mammalian Cells by Microinjection

Published on: December 4, 2010

15.9K

Assessing mRNA nuclear export in mammalian cells by microinjection

Eliza S Lee1, Alexander F Palazzo1

  • 1Department of Biochemistry, University of Toronto, Toronto, ON, Canada.

Methods (San Diego, Calif.)
|June 5, 2017
PubMed

Insights

This study details a method for measuring messenger RNA (mRNA) nuclear export rates in mammalian cells. Researchers can track mRNA movement after microinjecting DNA plasmids, providing insights into gene expression.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Nuclear export of messenger RNAs (mRNAs) is crucial for eukaryotic gene expression.
  • The mechanisms governing mRNA export are not fully understood.
  • Monitoring mRNA export is essential for studying gene regulation.

Purpose of the Study:

  • To describe a practical method for assessing mRNA nuclear export rates in mammalian cells.
  • To provide a protocol for setting up microscopy equipment for cell microinjections.
  • To enable researchers to quantify the nuclear export rate of specific mRNAs.

Main Methods:

  • Microinjection of DNA plasmids into the nucleus of mammalian tissue culture cells.
  • Microscopy setup utilizing a micromanipulator for precise cell microinjections.
  • Monitoring the spatial distribution of transcribed mRNA over time to determine export rates.

Main Results:

  • A detailed protocol for performing nuclear mRNA export assays is presented.
  • The method allows for the determination of nuclear export rates for individual mRNAs.
  • The setup facilitates efficient and accurate monitoring of mRNA export dynamics.

Conclusions:

  • The described microinjection and microscopy technique offers a straightforward approach to study mRNA nuclear export.
  • This method provides quantitative data on mRNA export rates, aiding in the understanding of gene expression regulation.
  • The protocol is valuable for researchers investigating mRNA transport and its role in cellular function.
Keywords:
Fluorescent in situ hybridizationGene expressionMammalian tissue culture cellsMicroinjectionmRNA nuclear export kinetics

More Related Videos

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells08:53

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells

Published on: May 16, 2017

9.2K
Analysis of Spliceosomal snRNA Localization in Human Hela Cells Using Microinjection07:35

Analysis of Spliceosomal snRNA Localization in Human Hela Cells Using Microinjection

Published on: August 6, 2019

6.5K

Related Concept Videos

Nuclear Export01:42

Nuclear Export

The nucleus restricts several proteins within and allows others to pass. The restricted proteins possess a nuclear retention sequence or NRS, anchoring them to the nuclear lamins and preventing their transport to the cytosol. The non-restricted proteins, after their synthesis, are transported to their site of action, such as the cytosol or other organelles, with the help of nuclear export signals or NES.
NES are of three types- the canonical 10-residue long leucine-rich signal and other...
5.1K
Nuclear Export of mRNA02:31

Nuclear Export of mRNA

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...
9.0K
Nuclear Export of mRNA02:31

Nuclear Export of mRNA

5.6K
Regulated mRNA Transport02:22

Regulated mRNA Transport

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...
7.1K
Nuclear Protein Sorting01:34

Nuclear Protein Sorting

Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
6.5K
Directionality of Nuclear Transport01:42

Directionality of Nuclear Transport

Ras-related nuclear protein or Ran is a small G protein that cycles between its GTP and GDP bound states. Ran specific regulators, a Ran GTPase Activating Protein or RanGAP present in the cytosol and a Ran guanine nucleotide exchange factor or RanGEF present inside the nucleus regulate GTP/GDP exchange. A high concentration of GTP inside the cells, in addition to this asymmetric distribution of  Ran-specific regulators, leads to a higher RanGTP concentration inside the nucleus. This...
4.7K