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Videos de Conceptos Relacionados

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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CRISPR01:59

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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CRISPR and crRNAs02:53

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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RNA Editing02:23

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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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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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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Un sistema dividido CRISPR/Cas13b para la regulación y edición del ARN condicional

Ying Xu1, Na Tian1, Huaxia Shi1

  • 1Department of Chemistry, Case Western Reserve University, 2080 Adelbert Road, Cleveland, Ohio 44106, United States.

Journal of the American Chemical Society
|February 22, 2023
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores desarrollaron un nuevo sistema CRISPR/Cas13b controlado por ácido abscísico (ABA) y luz. Este sistema regula con precisión los niveles y las modificaciones del ARN, avanzando las herramientas de investigación del ARN.

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Área de la Ciencia:

  • Biología molecular
  • Biotecnología
  • Biología del ARN

Sus antecedentes:

  • El sistema CRISPR/Cas13b es una herramienta poderosa para los estudios de ARN.
  • El control preciso de la actividad de Cas13b es crucial para comprender las funciones del ARN.
  • Las herramientas existentes a menudo interfieren con las actividades nativas de ARN.

Objetivo del estudio:

  • Diseñar un sistema dividido de Cas13b para la regulación condicional del ARN.
  • Desarrollar un sistema inducible por ABA para la manipulación precisa del ARN.
  • Para explorar el control basado en la luz de la deposición de la modificación del ARN.

Principales métodos:

  • Diseño de un sistema dividido CRISPR/Cas13b activado por el ácido abscísico (ABA).
  • Desarrollo de una división dCas13b inducible por ABA para la deposición m6A.
  • Utilizando un derivado ABA fotoactivado para el control mediado por la luz.
  • Demostrando la desregulación del ARN dependiente de la dosis y el tiempo.

Principales resultados:

  • Activación y desactivación condicionales de la actividad de Cas13b/dCas13b mediante ABA.
  • Deposición controlada temporalmente de m6A en sitios específicos de ARN.
  • Modulación inducida por la luz de las actividades del sistema dividido Cas13b/dCas13b.
  • Desregulación dependiente de la dosis y el tiempo de los ARN endógenos.

Conclusiones:

  • Los sistemas divididos Cas13b/dCas13b desarrollados ofrecen un control temporal y condicional preciso sobre la manipulación del ARN.
  • Estas plataformas amplían el conjunto de herramientas de regulación de CRISPR y ARN para entornos celulares nativos.
  • Los sistemas minimizan la interrupción funcional de los ARN endógenos, facilitando estudios avanzados de ARN.