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Updated: May 25, 2025
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Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
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A roadmap toward genome-wide CRISPR screening throughout the organism
Tess K Fallon1, Kristin A Knouse2
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Cell Genomics
|February 25, 2025
Summary
Genome-wide CRISPR screening in vivo offers powerful insights into mammalian physiology and disease. Recent advances in delivery, library design, and readout overcome challenges, enabling broader application of this genetic dissection tool.
Area of Science:
- Genetics
- Molecular Biology
- Mammalian Physiology
Background:
- Genome-wide CRISPR screening is a powerful tool for studying mammalian physiology and disease.
- Significant technical challenges hinder its in vivo application, including sgRNA delivery, library coverage, and phenotype selection.
Purpose of the Study:
- To review recent advances in CRISPR screening technologies for in vivo applications.
- To highlight solutions for overcoming technical challenges in delivering genome-wide libraries and selecting phenotypes.
- To discuss the potential for broader application of high-throughput genetic dissection in mammals.
Main Methods:
- Review of recent literature on CRISPR screening technologies.
- Analysis of advancements in single-guide RNA (sgRNA) delivery systems.
- Examination of novel library design strategies.
- Evaluation of improved phenotypic readout methods.
Main Results:
- Significant progress has been made in sgRNA delivery methods for in vivo CRISPR screening.
- Innovations in library design enhance coverage and efficiency.
- New phenotypic readout techniques improve the ability to select for desired outcomes.
- These advancements collectively address key technical hurdles.
Conclusions:
- Ongoing innovation in CRISPR screening components is paving the way for widespread in vivo application.
- Genome-wide CRISPR screening will become feasible in diverse mammalian cell types.
- This will enable unprecedented investigation into complex physiological and disease mechanisms.
