CRISPR-Mediated Library Screening of Gene-Knockout Cell Lines for Investigating Antiviral Innate Immunity.
Methods Mol Biol
; 2854: 51-60, 2025.
Article
em En
| MEDLINE
| ID: mdl-39192118
ABSTRACT
The application of CRISPR-mediated library screening has fundamentally transformed functional genomics by revealing the complexity of virus-host interactions. This protocol describes the use of CRISPR-mediated library screening to identify key functional genes regulating the innate immune response to PEDV infection. We detail a step-by-step process, starting from the design and construction of a customized CRISPR knockout library targeting genes involved in innate immunity to the effective delivery of these constructs into cells using lentiviral vectors. Subsequently, we outline the process of identifying functional genes postviral attack, including the use of next-generation sequencing (NGS), to analyze and identify knockout cells that exhibit altered responses to infection. This integrated approach provides researchers in immunology and virology with a resource and a robust framework for uncovering the genetic basis of host-pathogen interactions and the arsenal of the innate immune system against viral invasions.
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Base de dados:
MEDLINE
Assunto principal:
Biblioteca Gênica
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Técnicas de Inativação de Genes
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Sistemas CRISPR-Cas
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Imunidade Inata
Idioma:
En
Ano de publicação:
2025
Tipo de documento:
Article