简介：
Overview
This protocol describes a rapid and direct in vivo CRISPR/Cas9 screening methodology using ultrasound-guided in utero embryonic lentiviral injections. It allows researchers to assess the functions of several genes in the skin and oral cavity of immunocompetent mice.
Key Study Components
Area of Science
- Neuroscience
- Genetics
- Cancer Research
Background
- The technique enables probing of tumor suppressive functions of hundreds of genes.
- It is cost-effective compared to conventional knockout mouse models.
- The method is adaptable for studying gene expression in skin.
- It can be applied to various cancer types.
Purpose of Study
- To develop a methodology for simultaneous gene function assessment.
- To enhance understanding of gene roles in head and neck cancer.
- To provide a flexible tool for cancer research.
Methods Used
- Ultrasound-guided in utero embryonic lentiviral injections.
- Application of isoflurane and oxygen for anesthesia.
- Confirmation of pregnancy via hair removal cream application.
- Use of a targeted CRISPR library protocol.
Main Results
- Successful assessment of gene functions in vivo.
- Demonstrated cost-effectiveness of the method.
- Adaptability for different cancer types confirmed.
- Potential for broader applications in gene function studies.
Conclusions
- The protocol offers a rapid and efficient approach to gene function analysis.
- It opens new avenues for cancer research and gene studies.
- Future applications may extend beyond head and neck cancer.
What is the main advantage of this protocol?
It allows simultaneous assessment of multiple genes at a lower cost compared to traditional methods.
Can this method be used for other types of cancer?
Yes, the targeted CRISPR library protocol can be adapted for various cancer types.
How is anesthesia administered during the procedure?
Isoflurane and oxygen are supplied through a nosecone anesthesia tube.
What is the significance of using ultrasound guidance?
Ultrasound guidance enhances precision during the embryonic injections.
How do researchers confirm pregnancy in the mice?
Pregnancy is confirmed by applying hair removal cream to the abdomen.
What are the potential applications of this methodology?
It can be used for gene function studies in various tissues and cancer types.
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