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Related Concept Videos

Transcription01:10

Transcription

157.1K
Overview
Transcription is the process of synthesizing RNA from a DNA sequence by RNA polymerase. It is the first step in producing a protein from a gene sequence. Additionally, many other proteins and regulatory sequences are involved in the proper synthesis of messenger RNA (mRNA). Regulation of transcription is responsible for the differentiation of all the different types of cells and often for the proper cellular response to environmental signals.
Transcription Can Produce Different Kinds...
157.1K
Transcription Factors02:16

Transcription Factors

82.9K
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
82.9K
Eukaryotic Transcription Activators02:42

Eukaryotic Transcription Activators

12.9K
Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These...
12.9K
Transcription Attenuation in Prokaryotes02:42

Transcription Attenuation in Prokaryotes

18.5K
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure.  Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
18.5K
Master Transcription Regulators02:23

Master Transcription Regulators

7.8K
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a  complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
7.8K
5-Number Summary01:04

5-Number Summary

5.7K
In a dataset, the 5-number summary includes the minimum data value, the data value of the first quartile, the median data value or data value of the second quartile, the data value of the third quartile, and the maximum data value. These 5 data values can be visualized as a box and whisker plot.
In a box plot, the minimum and maximum data values represent the lower and upper whiskers in the graph, and the median is designated as the center of the box in the chart. The first quartile and third...
5.7K

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Related Experiment Video

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Quantification of the Abundance and Charging Levels of Transfer RNAs in Escherichia coli
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Very Low Abundance Single-Cell Transcript Quantification with 5-Plex ddPCRTM Assays.

George Karlin-Neumann1, Bin Zhang2, Claudia Litterst2

  • 1Digital Biology Center, Bio-Rad Laboratories, Pleasanton, CA, USA. George_karlin-neumann@bio-rad.com.

Methods in Molecular Biology (Clifton, N.J.)
|May 3, 2018
PubMed
Summary

This study presents a rapid 1-day workflow for single-cell gene expression analysis. It combines cell sorting with ultrahigh-sensitivity digital PCR for precise quantification of gene expression changes in individual cells.

Keywords:
Assay multiplexingCell-sortingDigital PCRDroplet digital PCRGene expressionRNA quantificationRadial multiplexingSingle-cellTranscriptsdPCRddPCR

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Area of Science:

  • Molecular Biology
  • Genomics
  • Cell Biology

Background:

  • Gene expression studies are crucial for understanding cellular phenotypes and responses.
  • Current methods like PCR and next-generation sequencing allow gene-focused or whole-transcriptome analysis.
  • Single-cell analysis, enabled by cell sorting, enhances resolution and sensitivity in gene expression profiling.

Purpose of the Study:

  • To present a rapid, efficient 1-day workflow for single-cell gene expression analysis.
  • To enable quantitative tracking of gene expression changes in individual cells.
  • To utilize a small laboratory cell-sorter and multiplexed digital PCR.

Main Methods:

  • Single-cell sorting using a compact laboratory cell-sorter.
  • Ultrahigh sensitivity, multiplexed digital PCR for gene expression quantification.
  • Workflow optimized for speed and efficiency within a 1-day timeframe.

Main Results:

  • Successful implementation of a 1-day workflow for single-cell analysis.
  • Quantitative tracking of expression levels for 5-10 genes per single cell.
  • Demonstrated high sensitivity and resolution in detecting gene expression changes.

Conclusions:

  • The presented workflow offers a quick and efficient method for single-cell gene expression profiling.
  • This approach strengthens inferences from observed expression levels and changes.
  • It provides a valuable tool for detailed molecular analysis of cellular heterogeneity.