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Ribozymes02:47

Ribozymes

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The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Riboswitches01:56

Riboswitches

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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.
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Bacterial RNA Polymerase00:43

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Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
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RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
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Ribozyme-Spherical Nucleic Acids.

Jessica L Rouge1,2, Timothy L Sita3,2,4, Liangliang Hao3,2

  • 1Department of Chemistry, Northwestern University, Evanston, Illinois 60208, United States.

Journal of the American Chemical Society
|August 15, 2015
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel spherical nucleic acid (SNA) architecture to stabilize ribozymes, enhancing their therapeutic potential. This new ribozyme-SNA effectively targets and reduces MGMT in glioblastoma cells, improving chemotherapy sensitivity.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • RNA Therapeutics

Background:

  • Ribozymes are RNA molecules with catalytic activity, capable of cleaving specific mRNA sequences.
  • Current ribozyme therapies face limitations due to large size and structural instability, hindering efficacy compared to siRNA and miRNA.
  • O(6)-methylguanine-DNA methyltransferase (MGMT) is a key protein in chemotherapeutic resistance, particularly in glioblastoma multiforme (GBM).

Purpose of the Study:

  • To develop a novel synthetic strategy for stabilizing ribozymes using spherical nucleic acid (SNA) architecture.
  • To characterize the therapeutic potential of ribozyme-SNA constructs for targeting MGMT in cancer cells.
  • To evaluate the efficacy of ribozyme-SNA in promoting apoptosis and sensitizing GBM cells to chemotherapy.

Main Methods:

  • Synthesis of ribozyme-SNA conjugates utilizing spherical nucleic acid architecture.
  • Characterization of ribozyme-SNA stability and cellular uptake.
  • Assessment of MGMT mRNA cleavage and protein knockdown in GBM cells.
  • Evaluation of GBM cell sensitization to therapy-induced apoptosis.

Main Results:

  • The novel ribozyme-SNA architecture successfully stabilized ribozymes for cellular delivery and function.
  • Direct cleavage of full-length MGMT mRNA was observed, leading to significant MGMT protein knockdown.
  • GBM cells treated with ribozyme-SNA showed increased sensitization to therapy-mediated apoptosis.
  • Effective delivery and function were achieved independently of traditional transfection agents.

Conclusions:

  • The spherical nucleic acid (SNA) architecture offers a promising strategy for stabilizing ribozymes and enhancing their therapeutic delivery.
  • Ribozyme-SNA conjugates represent a novel and effective approach for targeting MGMT in glioblastoma, potentially overcoming therapeutic resistance.
  • This innovative chemical architecture holds significant promise for advancing RNA-based therapeutics for solid tumors.