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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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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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CRISPR01:59

CRISPR

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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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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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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Conservative Site-specific Recombination and Phase Variation02:53

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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.
The recognition sites for Cre recombinase called LoxP...
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Diversos sistemas CRISPR-Cas codificados por virus incluyen editores de genoma simplificados

Basem Al-Shayeb1, Petr Skopintsev2, Katarzyna M Soczek2

  • 1Department of Plant and Microbial Biology, University of California, Berkeley, CA, USA; Innovative Genomics Institute, University of California, Berkeley, CA, USA; Department of Earth and Planetary Science, University of California, Berkeley, CA, USA; Department of Environmental Science, Policy and Management, University of California, Berkeley, CA, USA; University of Melbourne, Melbourne, Australia; Department of Chemistry, University of California, Berkeley, CA, USA; MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA; Gladstone Institutes, University of California, San Francisco, CA, USA.

Cell
|November 24, 2022
PubMed
Resumen

Los sistemas CRISPR-Cas, típicamente encontrados en microbios, también se descubren en bacteriófagos como herramientas antivirales compactas. Estos sistemas codificados por fagos, incluidas las nuevas nucleasas Casλ, muestran potencial para la edición del genoma en plantas y células humanas.

Palabras clave:
CRISPR y sus derivadosCRISPR-Cas también incluye:anti-viralesla enzimaEdición del genomaEdición del genomaMetagenomíael fagoLa estructuraherramienta

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Área de la Ciencia:

  • Microbiología
  • Biología molecular
  • La genética

Sus antecedentes:

  • Los sistemas CRISPR-Cas son mecanismos de defensa microbiana contra los virus.
  • Estos sistemas utilizan vías guiadas por ARN para la inmunidad adaptativa.

Objetivo del estudio:

  • Investigar la presencia y diversidad de los sistemas CRISPR-Cas en los bacteriófagos.
  • Caracterizar nuevas enzimas CRISPR-Cas, en particular de los bacteriófagos, para aplicaciones de edición del genoma.

Principales métodos:

  • Metagenómica resuelta por el genoma para identificar sistemas CRISPR codificados por fagos.
  • Microscopía criolectrónica para determinar la estructura de los complejos Casλ-crRNA-ADN.
  • Ensayos funcionales en células de mamíferos, Arabidopsis y trigo para evaluar las capacidades de edición del genoma.

Principales resultados:

  • Se identificaron sistemas CRISPR-Cas en diversos bacteriófagos, que funcionan como sistemas antivirales compactos.
  • Se descubrieron nuevas proteínas similares a Cas9 y 44 familias relacionadas con los sistemas CRISPR-Cas de tipo V, incluida la nucleasa Casλ.
  • La estructura Casλ-crRNA-DNA reveló una arquitectura única que permite la edición del genoma en varios tipos de células.

Conclusiones:

  • Los bacteriófagos representan una nueva fuente de diversos sistemas CRISPR-Cas.
  • Las enzimas CRISPR-Cas derivadas de fagos, como Casλ, ofrecen herramientas prometedoras para la edición del genoma en plantas y células humanas.