Tutorial: design and execution of CRISPR in vivo screens

Braun, Christian J.; Adames, Andres Carbonell; Saur, Dieter und Rad, Roland (2022): Tutorial: design and execution of CRISPR in vivo screens. In: Nature Protocols, Bd. 17, Nr. 9: S. 1903-1925

Abstract

This new tutorial from Braun and colleagues details how to design and execute in vivo CRISPR screens in mice to investigate a variety of phenotypes. Here we provide a detailed tutorial on CRISPR in vivo screening. Using the mouse as the model organism, we introduce a range of CRISPR tools and applications, delineate general considerations for 'transplantation-based' or 'direct in vivo' screening design, and provide details on technical execution, sequencing readouts, computational analyses and data interpretation. In vivo screens face unique pitfalls and limitations, such as delivery issues or library bottlenecking, which must be counteracted to avoid screening failure or flawed conclusions. A broad variety of in vivo phenotypes can be interrogated such as organ development, hematopoietic lineage decision and evolutionary licensing in oncogenesis. We describe experimental strategies to address various biological questions and provide an outlook on emerging CRISPR applications, such as genetic interaction screening. These technological advances create potent new opportunities to dissect the molecular underpinnings of complex organismal phenotypes.

• Dokumententyp: Zeitschriftenartikel
• Fakultät: Medizin
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• ISSN: 1754-2189
• Sprache: Englisch
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 329628492
• DFG: Gefördert durch die Deutsche Forschungsgemeinschaft (DFG) - 395357507