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Related Concept Videos

Alternative RNA Splicing02:18

Alternative RNA Splicing

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
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What is Gene Expression?01:36

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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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In eukaryotic cells, nascent mRNA transcripts need to undergo many post-transcriptional modifications to reach the cell cytoplasm and translate into functional proteins. For a long time, transcription and pre-mRNA processing were considered two independent events that occur sequentially in the cell. However, it has now been well established that transcription and pre-mRNA processing are two simultaneous processes that are precisely regulated inside the cell.
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Regulation of Expression at Multiple Steps01:23

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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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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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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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Using the E1A Minigene Tool to Study mRNA Splicing Changes
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Cell-specific regulation of gene expression using splicing-dependent frameshifting.

Jonathan P Ling1,2,3, Alexei M Bygrave4, Clayton P Santiago4

  • 1Departments of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA. jling@jhu.edu.

Nature Communications
|October 1, 2022
PubMed
Summary
This summary is machine-generated.

Scientists developed a new splicing-based gene delivery method called Splicing-Linked Expression Design (SLED). This approach precisely controls gene expression in specific cells, overcoming key challenges in genetic research and therapy.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Neuroscience

Background:

  • Cell-specific gene delivery is crucial but technically difficult.
  • Current methods often face limitations in precision and selectivity.
  • The need for robust tools to control gene expression in targeted cell populations is high.

Purpose of the Study:

  • To develop a novel splicing-based method for precise cell-specific gene expression control.
  • To overcome the tradeoff between promoter strength and selectivity in gene delivery.
  • To demonstrate the versatility and applicability of the new method in various research and therapeutic contexts.

Main Methods:

  • Developed Splicing-Linked Expression Design (SLED), a method using frameshifting alternative exons to control gene expression.
  • Analyzed RNA sequencing datasets to identify cell-specific exons based on specificity, conservation, and intron length.
  • Combined SLED with existing techniques like minipromoters and viral capsids (e.g., AAV).
  • Validated SLED vectors for delivering reporters and indicators in vivo and targeting disease-related genes.

Main Results:

  • SLED enables precise gene expression control by decoupling promoter strength from selectivity.
  • AAV-packaged SLED vectors successfully targeted specific neuronal subtypes in vivo.
  • Demonstrated gene therapy potential by creating SLED vectors for PRPH2 and SF3B1 mutations.
  • SLED technology offers flexibility for diverse research applications.

Conclusions:

  • Splicing-Linked Expression Design (SLED) provides a powerful and flexible platform for cell-specific gene delivery.
  • This method enhances precision and overcomes limitations of existing gene expression control strategies.
  • SLED holds significant promise for advancing basic research and developing novel gene therapies.