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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.
Ribozymes can...
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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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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.
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Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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Overview
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.
RNA...
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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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Elucidando los principios de diseño para la Especificidad de Tejido Habilitada por Ribozima (RETS) para permitir una

Max M Combest1, Josh Conlin1, Vivia Van De Mark2

  • 1Colorado State University Department of Biology.

bioRxiv : the preprint server for biology
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Resumen

Desarrollamos Ribozyme Enabled Tissue Specificity (RETS) para controlar la expresión transgénica en las plantas sin necesidad de promotores conocidos. Este método utiliza ribozimas para lograr una expresión génica precisa y específica para los biosensores y la ingeniería de cultivos.

Palabras clave:
Ciencias Biológicas AplicadasLas ribozimasExpresión específica del tejidoBiosensor de expresióndesarrollo de plantasBiología sintética vegetal

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Área de la Ciencia:

  • Biología vegetal
  • Biología sintética
  • Genética molecular

Sus antecedentes:

  • La expresión transgénica específica del tejido es crucial para los estudios biológicos y la ingeniería de organismos.
  • La identificación de promotores adecuados para una expresión precisa es un desafío, especialmente en las plantas debido a los largos plazos de creación de prototipos.
  • Los métodos existentes tienen limitaciones en el estudio de la expresión génica nativa y la ingeniería de fenotipos de plantas.

Objetivo del estudio:

  • Introducir una nueva estrategia, Ribozyme Enabled Tissue Specificity (RETS), para lograr la expresión transgénica específica del tejido en las plantas.
  • Permitir un control preciso de la expresión transgénica sin depender de promotores caracterizados.
  • Demostrar la utilidad de RETS para crear biosensores y características de ingeniería de plantas.

Principales métodos:

  • Desarrolló RETS, una estrategia que utiliza ribozimas autoesplicantes divididas (basadas en un intrón del grupo I de Tetrahymena thermophila).
  • Datos transcriptómicos aprovechados para guiar el diseño de la reconstitución condicional de ARNm.
  • Características de diseño optimizadas para la flexibilidad transgénica, la expresión mejorada y la evasión de la interferencia de ARN.

Principales resultados:

  • Se ha demostrado la expresión transgénica específica del tejido y dependiente de la dosis en Arabidopsis thaliana utilizando RETS.
  • Mostró la creación de biosensores codificados genéticamente para el estudio de la expresión génica espacio-temporal en plantas.
  • Ilustró la ingeniería de cambios específicos del tejido en el tamaño del órgano, demostrando el control fenotípico.

Conclusiones:

  • RETS ofrece un nuevo enfoque para estudiar los patrones de expresión génica nativa utilizando imágenes no destructivas, superando las limitaciones de las técnicas actuales.
  • El control espacio-temporal de la expresión transgénica a través de RETS permite la ingeniería de precisión de los fenotipos de las plantas.
  • Esta tecnología facilita la mejora de los cultivos sin los inconvenientes de la expresión constitutiva, allanando el camino para mejores aplicaciones agrícolas.