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Cell Specific Gene Expression

Cell Specific Gene Expression

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.

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Using Fluorescence Activated Cell Sorting to Examine Cell-Type-Specific Gene Expression in Rat Brain Tissue

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Cell Type-Specific Gene Expression Profiling Using Fluorescence-Activated Nuclear Sorting.

Daniel Slane¹, Martin Bayer²

¹Department of Cell Biology, Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076, Tuebingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|June 18, 2017

Summary

This summary is machine-generated.

This study presents a new protocol for Fluorescence-Activated Cell Sorting (FACS) analysis of plant nuclei, enabling transcriptional profiling of previously inaccessible tissues. This method successfully mapped the early Arabidopsis thaliana embryo transcriptome.

Keywords:

Cell type FACS FANS Microarray Nuclear transcriptome RNA amplification mRNA

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Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

Area of Science:

Plant molecular biology
Genomics
Cell biology

Background:

Fluorescence-activated cell sorting (FACS) is crucial for analyzing cell-specific molecular profiles.
Current FACS methods in plants are limited to easily dissociable tissues like roots and shoots.
Many plant tissues are recalcitrant to cell separation, hindering comprehensive analysis.

Purpose of the Study:

To develop a detailed protocol for generating transcriptional profiles from fluorescently labeled plant nuclei.
To overcome limitations of current FACS methods for recalcitrant plant tissues.
To enable transcriptomic analysis of previously inaccessible plant cell types.

Main Methods:

A novel protocol for generating transcriptional profiles from fluorescently labeled nuclei.
Application of the protocol to early Arabidopsis thaliana embryo tissue.
Cell wall digestion and nuclear isolation techniques adapted for recalcitrant tissues.

Main Results:

Successful generation of transcriptional profiles from isolated nuclei.
Demonstration of the protocol's efficacy on early Arabidopsis thaliana embryos.
Creation of a detailed transcriptional map of the early embryo.

Conclusions:

The developed protocol significantly expands the applicability of FACS in plant research.
This method allows for transcriptomic analysis of plant nuclei from recalcitrant tissues.
The transcriptional map of the early Arabidopsis thaliana embryo provides valuable insights into early development.