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Related Concept Videos

MicroRNAs01:22

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MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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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...
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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.
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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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miRNA modules for precise, tunable control of gene expression.

Rongrong Du1,2,3, Michael J Flynn1,2,3, Monique Honsa1,2

  • 1Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.

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Researchers developed DIMMERs, synthetic miRNA-based gene expression regulators. These circuits offer precise, tunable control of transgenes, enabling consistent protein levels across various cell types for research and therapy.

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Area of Science:

  • Synthetic biology
  • Molecular and cell biology
  • Biotechnology

Background:

  • Precise control of transgene expression is crucial for cellular research and therapeutic applications.
  • Current gene delivery methods lack quantitative control and are often inefficient.
  • Existing compensation mechanisms have limitations in tunability and complexity.

Purpose of the Study:

  • To develop a novel toolkit of synthetic gene expression regulators for precise and tunable transgene control.
  • To achieve dosage-invariant protein expression across varying genetic contexts.
  • To enable independent control of multiple genes within the same cell.

Main Methods:

  • Design and implementation of compact, synthetic microRNA (miRNA)-based circuit modules (DIMMERs).
  • Utilizing multivalent miRNA regulatory interactions within an incoherent feed-forward loop architecture.
  • Development of a heuristic miRNA design algorithm for orthogonal circuit variants.

Main Results:

  • DIMMERs achieve nearly uniform protein expression over a wide range of gene dosages and transcription rates.
  • Demonstrated precise, tunable, and dosage-invariant control of transgenes in diverse cell types.
  • Successfully applied DIMMERs to enhance CRISPR imaging and super-resolution EGFR receptor imaging.

Conclusions:

  • The DIMMERs toolkit provides a powerful solution for precise and tunable gene expression control.
  • These regulators are portable across different cell types and applicable to various biotechnology fields.
  • The developed system advances gene therapy and fundamental biological research through improved transgene expression management.