Video Experimental Relacionado
Updated: Feb 5, 2026
08:20
Genome Engineering of Primary Human B Cells Using CRISPR/Cas9
Published on: November 3, 2020
7.0K
Nucleasa CRISPR-Cas9 diseñada con espacio de orientación expandido
Hiroshi Nishimasu1, Xi Shi2,3, Soh Ishiguro4,5,6
1Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. nisimasu@bs.s.u-tokyo.ac.jp nureki@bs.s.u-tokyo.ac.jp.
Science (New York, N.Y.)
|September 1, 2018
Resumen
Los científicos diseñaron una variante de la enzima Cas9 (SpCas9-NG) para reconocer motivos adyacentes de protospacer NG relajados (PAM). Esto expande las capacidades de edición del genoma al apuntar a los loci genómicos previamente inaccesibles en las células humanas.
Área de la Ciencia:
- Biología molecular
- La genómica
- La bioquímica
Sus antecedentes:
- La enzima Cas9 es una herramienta clave en la edición del genoma, que permite la escisión del ADN dirigido.
- La enzima Streptococcus pyogenes Cas9 (SpCas9) comúnmente utilizada está limitada por su requisito para un motivo adyacente de protospacer NGG específico (PAM).
- Esta especificidad NGG PAM restringe el rango de sitios genómicos que pueden ser objeto de edición.
Objetivo del estudio:
- Para diseñar una variante SpCas9 con capacidades de reconocimiento PAM relajadas.
- Para superar las limitaciones impuestas por el estricto requisito NGG PAM del tipo salvaje SpCas9.
- Ampliar el alcance de los loci genómicos seleccionables para la edición del genoma basada en CRISPR.
Principales métodos:
- Ingeniería racional de la enzima SpCas9 para crear una variante (SpCas9-NG) con especificidad PAM alterada.
- Cristalografía de rayos X para determinar la base estructural del reconocimiento de NG PAM por SpCas9-NG.
- Ensayos funcionales en células humanas para evaluar la capacidad de SpCas9-NG para inducir indels en los sitios de NG PAM.
- Fusión de SpCas9-NG con desaminasa citidina inducida por activación (AID) para probar las capacidades de edición de bases en los sitios NG PAM.
Principales resultados:
- Se ha diseñado con éxito una variante SpCas9-NG capaz de reconocer NG PAM relajados.
- La estructura cristalina reveló que SpCas9-NG acomoda NG PAM a través de interacciones específicas de base alteradas.
- SpCas9-NG demostró eficacia en la inducción de inserciones/deleciones (indels) en sitios endógenos de NG PAM en células humanas.
- SpCas9-NG fusionado con AID facilitó las conversiones dirigidas de C a T en los sitios de NG PAM en células humanas.
Conclusiones:
- SpCas9-NG representa un avance significativo en la tecnología de edición del genoma basada en Cas9.
- El SpCas9-NG diseñado amplía el rango de orientación, permitiendo la edición en loci genómicos previamente inaccesibles.
- La fusión SpCas9-NG con AID ofrece una herramienta versátil para la edición precisa de bases en los sitios NG PAM, mejorando las aplicaciones de ingeniería genómica.
Videos de Conceptos Relacionados
01:59CRISPR
57.9K
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...
57.9K
02:53CRISPR and crRNAs
19.1K
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...
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...
19.1K
00:49What is Genetic Engineering?
80.3K
Overview
80.3K
01:28CRISPR/Cas9 Genome Editing
1.9K
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...
1.9K
01:25Space Trusses
1.3K
A space truss is a three-dimensional counterpart of a planar truss. These structures consist of members connected at their ends, often utilizing ball-and-socket joints to create a stable and versatile framework. The space truss is widely used in various construction projects due to its adaptability and capacity to withstand complex loads.
At the core of a space truss lies the fundamental unit known as the tetrahedron. This structure is composed of six members that form a three-dimensional shape...
At the core of a space truss lies the fundamental unit known as the tetrahedron. This structure is composed of six members that form a three-dimensional shape...
1.3K
01:27State Space Representation
583
The frequency-domain technique, commonly used in analyzing and designing feedback control systems, is effective for linear, time-invariant systems. However, it falls short when dealing with nonlinear, time-varying, and multiple-input multiple-output systems. The time-domain or state-space approach addresses these limitations by utilizing state variables to construct simultaneous, first-order differential equations, known as state equations, for an nth-order system.
Consider an RLC circuit, a...
Consider an RLC circuit, a...
583