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Related Concept Videos

Regulated Protein Degradation02:58

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
Regulated Protein Degradation02:58

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
Riboswitches01:56

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order to...
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
Covalently Linked Protein Regulators02:04

Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.

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Reliably Engineering and Controlling Stable Optogenetic Gene Circuits in Mammalian Cells
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Published on: July 6, 2021

A genetic bistable switch utilizing nonlinear protein degradation.

Daniel Huang1, William J Holtz, Michel M Maharbiz

  • 1Department of Electrical Engineering and Computer Science, University of California, 656 Sutardja Dai Hall,Berkeley, Berkeley, CA, 94720, USA. maharbiz@eecs.berkeley.edu.

Journal of Biological Engineering
|July 11, 2012
PubMed
Summary
This summary is machine-generated.

Engineered bistable switches in bacteria can now use both transcriptional and enzymatic parts. This hybrid system is tunable, retains states through cell division, and offers a more robust circuit design.

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Area of Science:

  • Synthetic biology
  • Genetic circuit design
  • Prokaryotic systems

Background:

  • Bistability is crucial for state switching and retention in biological and engineered systems.
  • Previous synthetic bistable switches in prokaryotes primarily used transcriptional elements.
  • Hybrid systems combining transcriptional and enzymatic components offer expanded parameter ranges.

Purpose of the Study:

  • To demonstrate a tunable family of hybrid bistable switches in E. coli.
  • To integrate transcriptional and enzymatic components for enhanced bistability.
  • To explore the tunability and robustness of such hybrid circuits.

Main Methods:

  • Constructed a synthetic bistable switch using lambda repressor (CI) and Mesoplasma florum Lon protease (mf-Lon).
  • Utilized two linked positive feedback loops, one transcriptional and one enzymatic.
  • Experimentally validated bistable behavior and state retention over 40 cell divisions in E. coli.

Main Results:

  • The hybrid bistable switch successfully retained its state without an input signal across 40 cell division cycles.
  • Switching speed was found to be tunable by altering the expression rate of mf-Lon.
  • Demonstrated a functional hybrid bistable switch in E. coli.

Conclusions:

  • This study presents the first use of dynamic expression of an orthogonal, heterologous protease to tune a nonlinear protein degradation circuit.
  • The developed hybrid switch offers a potentially more robust and tunable topology for prokaryotic synthetic biology applications.
  • Highlights the advantages of integrating enzymatic components into transcriptional circuits for enhanced control and performance.