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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
Published on: June 16, 2017
CRISPR/Cas9-The ultimate weapon to battle infectious diseases?
M Doerflinger1,2, W Forsyth1, G Ebert1,2
1Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
Abstract:
Infectious diseases are a leading cause of death worldwide. Novel therapeutics are urgently required to treat multidrug-resistant organisms such as Mycobacterium tuberculosis and to mitigate morbidity and mortality caused by acute infections such as malaria and dengue fever virus as well as chronic infections such as human immunodeficiency virus-1 and hepatitis B virus. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, which has revolutionized biomedical research, holds great promise for the identification and validation of novel drug targets. Since its discovery as an adaptive immune system in prokaryotes, the CRISPR/Cas9 system has been developed into a multi-faceted genetic modification tool, which can now be used to induce gene deletions or specific gene insertions, such as conditional alleles or endogenous reporters in virtually any organism. The generation of CRISPR/Cas9 libraries that can be used to perform phenotypic whole genome screens provides an important new tool that will aid in the identification of critical host factors involved in the pathogenesis of infectious diseases. In this review, we will discuss the development and recent applications of the CRISPR/Cas9 system used to identify novel regulators, which might become important in the fight against infectious diseases.
Insights
The CRISPR/Cas9 system revolutionizes infectious disease research by enabling genetic modifications for identifying new drug targets. This technology aids in combating drug-resistant bacteria and viruses like tuberculosis and HIV.
Area of Science:
- Microbiology
- Genetics
- Immunology
Background:
- Infectious diseases pose a significant global health threat, necessitating novel therapeutics.
- Multidrug-resistant organisms (e.g., Mycobacterium tuberculosis) and viruses (e.g., HIV-1, Hepatitis B) require innovative treatment strategies.
- The CRISPR/Cas9 system, initially an prokaryotic immune mechanism, is now a versatile genetic engineering tool.
Purpose of the Study:
- To review the development and applications of the CRISPR/Cas9 system in infectious disease research.
- To highlight CRISPR/Cas9's role in identifying novel drug targets and host factors.
- To discuss the potential of CRISPR/Cas9 in combating infectious diseases.
Main Methods:
- CRISPR/Cas9 system for genetic modification (deletions, insertions).
- Generation of CRISPR/Cas9 libraries for phenotypic whole-genome screens.
- Application of CRISPR/Cas9 in identifying host factors in infectious disease pathogenesis.
Main Results:
- CRISPR/Cas9 enables precise genetic modifications in various organisms.
- CRISPR/Cas9 libraries facilitate whole-genome screens for identifying critical host factors.
- The system aids in validating novel drug targets for infectious diseases.
Conclusions:
- The CRISPR/Cas9 system is a powerful tool for advancing infectious disease research.
- It accelerates the discovery of therapeutic targets against challenging pathogens.
- CRISPR/Cas9 holds significant promise for developing new strategies against infectious diseases.
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