Name: crispr-cas9 gene editing in action
Uploaded: 2019-03-11T09:21:22.000Z
Duration: 1 min 59 s
Channel: JoVE

K12 Standards

Tennessee Academic Standards

High School

Biology

links-among-engineering-technology-science-and-society

Links Among Engineering, Technology, Science, and Society

BIO1.ETS2.3

CRISPR

- [Instructor] El Sistema
CRISPR-Cas9 es una herramienta

que sirve para editar ADN. Las
siglas en inglés responden a:

Repeticiones Palindrómicas
Cortas Agrupadas y

Regularmente Interespaciadas

y a la Proteína 9 asociada al CRISPR.

Al observársele en las bacterias,

el CRISPR-Cas9 es un medio
de defensa contra los virus.

Cuando un ADN viral
extraño entra a la bacteria

es procesado en pequeños fragmentos,

los cuales se insertan en una zona

del genoma bacterial llamada Locus CRISPR.

Cuando se transcribe
la región, el producto

se asocia con pequeños
ARN llamados tracrARN,

los cuales ayudan a
orientar a la Proteína Cas9

y a los ARN hacia las moléculas.

Este último rompe la transcripción

y como resultado final
obtenemos varios complejos

consistentes en Proteína
Cas9, tracrARN y crARN,

derivados del ADN en el Locus.

El CRISPR ARN en estas
estructuras reconoce

y guía a la Cas9 al ADN viral,

el cual es descompuesto y destruido.

Los científicos obtienen CRISPR-Cas9

mediante la síntesis de
moléculas individuales de ARN

que imitan el tracrARN y el CRISPR ARN

para identificar genes sospechosos.

Por ejemplo, cuando dos de estos ARN

se introducen en células con Cas9

y ambos identifican al mismo gen,

se erradica una secuencia.

Y una vez que se remueve el objetivo

los extremos sueltos son reconectados

y se observan los efectos en las células.

Así logra modificarse
el Sistema CRISPR-Cas9

en un mecanismo bacterial
y puede aprovecharse

en una gran variedad de
técnicas de edición de genes.

Thurtle‐Schmidt, Deborah M. y Te‐Wen Lo. "Biología molecular a la vanguardia: una revisión sobre la edición genética CRISPR/CAS9 para estudiantes universitarios". Educación en Bioquímica y Biología Molecular 46, no. 2 (2018): 195–205. [<a href="https://doi.org/10.1002/bmb.21108" target="__blank">Fuente</a>]

Lander, Eric S. "Los héroes de CRISPR", Celda 164, núm. 1 (14 de enero de 2016): 18–28. [<a href="https://doi.org/10.1016/j.cell.2015.12.041" target="__blank">Fuente</a>]

crispr

Las tecnologías de edición del genoma permiten a los científicos modificar el ADN de un organismo mediante la adición, eliminación o reordenamiento de material genético en ubicaciones genómicas específicas. Este tipo de técnicas podrían usarse potencialmente para curar trastornos genéticos como la hemofilia y la anemia falciforme. Una herramienta de investigación de edición de ADN popular y ampliamente utilizada que podría conducir a curas seguras y eficaces para los trastornos genéticos es el sistema CRISPR-Cas9. CRISPR-Cas9 significa repeticiones palindrómicas cortas agrupadas regularmente inter está formada por unas endonucleasa Cas9 y un pequeño ARN que guía a Cas9 hacia el ADN objetivo. <h4>Origen</h4> Las secuencias CRISPR se observaron por primera vez en bacterias y luego se identificaron en arqueas. Los investigadores descubrieron que el sistema CRISPR-Cas9 sirve como defensa inmune adaptativa contra virus invasores. Muchas bacterias y la mayoría de las arqueas capturan secuencias cortas del ADN viral para crear una biblioteca de segmentos de ADN del virus o matrices CRISPR. Cuando los procariotas se vuelven a exponer al mismo virus o clase de virus, se utilizan matrices CRISPR para transcribir pequeños segmentos de ARN que ayudan a reconocer a los invasores virales y posteriormente a destruir el ADN viral con Cas9 o una endonucleasa similar. <h4>Usando la tecnología CRISPR-Cas9</h4> CRISPR-Cas9 se usa comúnmente en el laboratorio para eliminar ADN e insertar una nueva secuencia de ADN en su lugar. Para lograr esto, los investigadores primero deben crear un pequeño fragmento de ARN llamado ARN guía, con una secuencia corta llamada secuencia guía que se une a una secuencia objetivo específica en el ADN genómico. El ARN guía también puede asociarse con Cas9 (u otras endonucleasas como Cpf1). El ARN guía y la proteína Cas9 se administran a una célula de interés donde el ARN guía identifica la secuencia de ADN objetivo y Cas9 la escinde. Luego, la maquinaria de la célula repara las hebras rotas insertando o eliminando nucleótidos aleatorios, inactivando el gen objetivo. Alternativamente, se puede introducir una secuencia de ADN personalizada en la célula junto con el ARN guía y Cas9, que sirve como plantilla para la maquinaria de reparación y reemplaza la secuencia extirpada. Esta es una forma muy eficaz para que los investigadores "eliminen" un gen para estudiar sus efectos o reemplacen un gen mutado con una copia normal con la esperanza de curar una enfermedad. <h4>Consideraciones éticas y de viabilidad en humanos</h4> Como resultado de las importantes capacidades de modificación genética del sistema CRISPR-Cas9, ha habido un gran debate sobre su uso, especialmente en lo que respecta a la edición de embriones. Un científico chino afirmó recientemente haber creado bebés con genoma editado utilizando tecnología CRISPR para desactivar un gen implicado en la infección por VIH. Esto provocó una protesta mundial por parte de científicos preocupados por las consideraciones éticas y de seguridad del procedimiento. Muchos han calificado la medida como prematura y otros han expresado su preocupación por los efectos genómicos no deseados. Si bien la cantidad de posibles aplicaciones biotecnológicas para el sistema CRISPR-Cas9 es numerosa, es importante considerar los desafíos futuros que pueden surgir como resultado de su uso.

Las tecnologías de edición del genoma permiten a los científicos modificar el ADN de un organismo mediante la adición, eliminación o reorganización de material genético en un momento específico.

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KEY TERMS AND CONCEPTS

TÉRMINOS Y CONCEPTOS CLAVE

crispr-cas9 gene editing in action

CRISPR-Cas9 is a genome editing tool that lets scientists change DNA at a specific location. It can add, remove, or rearrange genetic material. Researchers study it because it may help treat genetic disorders such as hemophilia and sickle cell anemia.

CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats, and Cas9 is a CRISPR-associated protein 9. The basic system includes a Cas9 endonuclease, which is an enzyme that cuts DNA, and a small RNA that guides Cas9 to the target sequence. Together, they make CRISPR-Cas9 a precise research tool for DNA editing.

CRISPR sequences were first seen in bacteria and later found in archaea. Scientists discovered that this system acts like an adaptive immune defense against viruses. Many bacteria and most archaea store short pieces of viral DNA in CRISPR arrays. When the same virus returns, these RNA segments help identify it, and Cas9 or a similar endonuclease destroys the viral DNA.

In the lab, CRISPR-Cas9 is often used to remove DNA and replace it with a new sequence. Researchers first design a guide RNA, which contains a guide sequence that matches a target sequence on genomic DNA. The guide RNA and Cas9 are then placed in the cell of interest. The guide RNA finds the target, Cas9 cuts the DNA, and the cell repairs the break by adding or deleting random nucleotides, which can turn the gene off.

Scientists can also add a custom DNA template with the guide RNA and Cas9. The repair machinery can use that template to replace the removed sequence. This makes CRISPR useful for gene knockouts and for replacing a mutated gene with a normal copy in hopes of treating disease. Other endonucleases, such as Cpf1, can also work with guide RNA.

CRISPR-Cas9 has also raised major ethical and safety questions, especially about embryo editing. A Chinese scientist claimed to have used CRISPR to create genome-edited babies by disabling a gene linked to HIV infection. That claim led to global concern because many scientists thought the work was premature and worried about off-target genomic effects. CRISPR has many possible biotech uses, but future challenges still need careful attention.

CRISPR-Cas9 Gene Editing in Action

what is genetic engineering?

How Scientists Edit DNA for New Traits

Genetic engineering changes an organism’s DNA to add useful traits. Scientists use it in bacteria, plants, animals, and even viruses for research, medicine, agriculture, and industry. It can bring major benefits, but it also raises ethical concerns about humans and the food supply. This work is possible because DNA has the same basic structure and genetic code across life. That shared code lets scientists modify DNA in several ways. One method is CRISPR/Cas9, often called molecular scissors,...

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Creating Transgenic Plants and Animals

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in-vitro mutagenesis (gene knockouts)

Knockout Mice and Gene Replacement

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CRISPR-Cas9 is a genome editing tool that lets scientists change DNA at a specific location. It can add, remove, or rearrange genetic material. Researchers study it because it may help treat genetic disorders such as hemophilia and sickle cell anemia. CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats, and Cas9 is a CRISPR-associated protein 9. The basic system includes a Cas9 endonuclease, which is an enzyme that cuts DNA, and a small RNA that guides Cas9 to the...

gene therapy

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reproductive cloning

Cloning Animals with SCNT

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genomics

Genomics and DNA Sequencing

Genomics studies the full set of genetic material in an organism. In humans, that genome includes the DNA carried in 23 pairs of chromosomes in the nucleus, along with mitochondrial DNA. Genomics looks at both coding DNA and non-coding DNA, which helps scientists study how living things are built and how they differ. This field is useful in many areas of biology and medicine. Scientists use genomics to build phylogenetic trees, improve crop productivity and sustainability, help with criminal...

key terms and concepts

Learn how CRISPR-Cas9 edits DNA, uses guide RNA and Cas9, and why scientists debate its use in embryos and gene therapy.

CRISPR-Cas9 Gene Editing in Action | JoVE K12

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JoVE Core Biología da vida a la biología a través de una serie con más de 300 lecciones de vídeo animadas, concisas y fáciles de entender, que explican conceptos clave de la biología, incluyendo también más de 150 vídeos de científicos en acción, que muestran experimentos reales realizados en laboratorios actuales de investigación.

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Obtain, evaluate, and communicate information on how molecular biotechnology may be used in a variety of fields.
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EXPLANATION: The utilization of new technologies in any field of science is dependent on both economic and social factors. In addition to evaluating these factors, scientists must also consider long-term consequences that may not be initially apparent. The emphasis should be on the construction of a rationale argument that supports a position on the use of an application with ethical and social impact. Students should begin to appreciate the differences in ethical values that exist, and recognize that discussion of these values is imperative as knowledge and technology continue to advance, even when resolutions of differences can be rare. After investigation of a specific biotechnology application, students can write a position paper and/or participate in a classroom debate.

CRISPR-Cas9 Gene Editing in Action

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