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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...
Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form dimers that...
Cooperative Binding of Transcription Regulators02:13

Cooperative Binding of Transcription Regulators

Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form dimers that...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Transcriptional Regulation: Riboswitches01:23

Transcriptional Regulation: Riboswitches

Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...
Master Transcription Regulators02:23

Master Transcription Regulators

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...

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Identification of Functional Protein Regions Through Chimeric Protein Construction
11:39

Identification of Functional Protein Regions Through Chimeric Protein Construction

Published on: January 8, 2019

A modular strategy for engineering orthogonal chimeric RNA transcription regulators.

Melissa K Takahashi1, Julius B Lucks

  • 1School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853, USA.

Nucleic Acids Research
|June 14, 2013
PubMed
Summary

Researchers developed a modular strategy to create new orthogonal RNA transcription regulators. This advance enables the construction of more complex RNA genetic circuitry for synthetic biology applications.

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Area of Science:

  • Synthetic biology
  • Molecular biology
  • RNA biology

Background:

  • Antisense RNA transcription attenuators are crucial for synthetic biology, enabling signal integration and transcriptional cascades.
  • A major hurdle in developing advanced RNA genetic circuitry is the creation of orthogonal attenuators that function independently.

Purpose of the Study:

  • To develop a modular strategy for creating larger families of orthogonal antisense RNA transcription attenuators.
  • To engineer chimeric attenuators by fusing an engineered attenuator with natural antisense RNA translational regulators.

Main Methods:

  • Developed chimeric fusions between the pT181 transcriptional attenuator and five natural antisense RNA translational regulators.
  • Utilized in vivo gene expression assays in Escherichia coli for functional validation.
  • Performed mutagenesis to generate new chimeric attenuators and conducted orthogonality tests.

Main Results:

  • Successfully created chimeric attenuators by combining engineered and natural regulatory sequences.
  • Generated 11 new chimeric attenuators through mutagenesis.
  • Established a 7x7 matrix of mutually orthogonal regulators, demonstrating high functional independence.

Conclusions:

  • The modular strategy facilitates the engineering of orthogonal RNA transcription regulators.
  • The findings provide design principles for future RNA genetic circuitry development.
  • This work accelerates the creation of sophisticated RNA genetic circuits and advances non-coding RNA regulation understanding.