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JoVE Journal
Genetics
Manipulación CRISPR placentaria dirigida a ratones en vivo
Manipulación CRISPR placentaria dirigida a ratones en vivo
JoVE Journal
Genetics
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JoVE Journal Genetics
Mouse In Vivo Placental Targeted CRISPR Manipulation

Manipulación CRISPR placentaria dirigida a ratones en vivo

Full Text
3,856 Views
07:39 min
April 14, 2023

DOI: 10.3791/64760-v

Annemarie J. Carver1,2,3, Robert J. Taylor2,3, Hanna E. Stevens1,2,3,4

1Interdisciplinary Graduate Program in Genetics,University of Iowa, 2Department of Psychiatry, Carver College of Medicine,University of Iowa, 3Iowa Neuroscience Institute, Carver College of Medicine,University of Iowa, 4Hawk-IDDRC,University of Iowa

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol describes a time-specific method for manipulating critical developmental pathways in the mouse placenta using CRISPR. The technique involves the injection and electroporation of CRISPR plasmids into the placentas of pregnant mice at embryonic day 12.5.

Key Study Components

Area of Science

  • Neuroscience
  • Developmental Biology
  • Genetic Engineering

Background

  • Targeted manipulation of gene functions during pregnancy is crucial for understanding developmental processes.
  • Genetic modifications in the placenta are rare and challenging.
  • This method allows for a range of gene expression changes.
  • Mastery of the technique requires practice and persistence.

Purpose of Study

  • To elucidate specific gene functions during mid to late pregnancy.
  • To provide a reliable method for gene manipulation in the mouse placenta.
  • To advance the understanding of placental development and function.

Methods Used

  • Injection of CRISPR plasmids into mouse placentas.
  • Electroporation to facilitate plasmid uptake.
  • Timing of intervention at embryonic day 12.5.
  • Assessment of gene expression changes post-manipulation.

Main Results

  • Successful manipulation of gene expression in the mouse placenta.
  • Demonstrated potential for overexpression of specific genes.
  • Provided insights into the challenges and learning curve associated with the technique.
  • Highlighted the significance of time-specific interventions.

Conclusions

  • This method represents a significant advancement in placental research.
  • It opens new avenues for studying gene function during pregnancy.
  • Researchers are encouraged to persist through initial challenges.

Frequently Asked Questions

What is the significance of this protocol?
It allows for targeted manipulation of mouse placentas using CRISPR, aiding in the study of gene functions during pregnancy.
What challenges might researchers face?
The technique is challenging and may require time to master, with initial attempts possibly being unsuccessful.
At what embryonic stage is the intervention performed?
The intervention is performed on embryonic day 12.5.
What are the main advantages of this method?
It allows for a range of gene expression changes within the placenta, which is uncommon in existing techniques.
How can researchers improve their success with this technique?
Practice and persistence are key; researchers should not be discouraged by initial failures.

Aquí describimos un método específico en el tiempo para manipular eficazmente las vías críticas de desarrollo en la placenta del ratón in vivo. Esto se realiza mediante la inyección y electroporación de plásmidos CRISPR en las placentas de madres embarazadas en el día embrionario 12.5.

Este protocolo es significativo ya que permite la manipulación dirigida en tiempo específico de placentas de ratón utilizando CRISPR. Esto permitirá a los investigadores dilucidar funciones genéticas específicas durante la mitad y el final del embarazo. Las manipulaciones genéticas que causan sobreexpresión dentro de la placenta del ratón son muy poco comunes.

La principal ventaja de esta técnica es que permite una serie de cambios en la expresión génica dentro de la placenta. No se desanime si los términos preliminares que tiene esta técnica no tienen éxito. Como esta técnica es desafiante, requiere bastante tiempo para practicar para dominarla.

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