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Related Concept Videos

Combinatorial Gene Control02:33

Combinatorial Gene Control

Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
Cell Specific Gene Expression01:58

Cell Specific Gene Expression

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...
Cell Specific Gene Expression01:58

Cell Specific Gene Expression

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...
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
Constitutive and Regulated Gene Expression01:27

Constitutive and Regulated Gene Expression

Gene expression in prokaryotes is governed by constitutive and regulated systems, allowing cells to balance the production of essential proteins with adaptive responses to environmental changes.Constitutive Gene ExpressionConstitutive, or housekeeping, genes are continuously expressed as they encode proteins vital for fundamental cellular processes. These include enzymes for glycolysis, ribosomal components for protein synthesis, and proteins involved in DNA replication. Their constant...
Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...

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Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX
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pHUSH: a single vector system for conditional gene expression.

Daniel C Gray1, Klaus P Hoeflich, Li Peng

  • 1Department of Molecular Biology, Genentech Inc, South San Francisco, California, USA. Daniel.Gray@ucsf.edu

BMC Biotechnology
|September 28, 2007
PubMed
Summary
This summary is machine-generated.

We developed pHUSH, a single-vector system for inducible gene expression, overcoming limitations of multi-plasmid systems. This tool enables conditional RNAi or transgene expression in various models, advancing genetic engineering research.

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Mammalian Cell Engineering

Background:

  • Traditional gene expression studies face challenges with artefacts and multi-plasmid systems.
  • Existing methods require extensive engineering, causing delays and potential cell subpopulation bias.
  • A single-vector inducible system is needed to streamline research and improve accuracy.

Purpose of the Study:

  • To develop a novel, single-vector inducible expression system for regulated gene expression.
  • To overcome the limitations of multi-plasmid systems in conditional gene studies.
  • To facilitate RNA interference (RNAi) and transgene expression studies.

Main Methods:

  • Development of the pHUSH (plasmid-based inducible system) vector.
  • Integration of shRNA, miRNA, or cDNA expression cassettes.
  • Incorporation of Pol II and Pol III promoters with a tetracycline repressor and selectable marker.

Main Results:

  • pHUSH successfully enables Doxycycline-regulated RNAi or transgene expression.
  • The system utilizes a single viral vector for efficient delivery and expression.
  • Demonstrated successful application in vitro and in vivo, including conditional gene knockdown in a brain cancer model.

Conclusions:

  • pHUSH is a versatile single-vector system for conditional genetic engineering.
  • The system supports Doxycycline-regulated expression of RNAi or transgenes.
  • pHUSH shows broad applicability in mammalian cell research and in vivo models.