Tutorial: design and execution of CRISPR in vivo screens

Autor:	Christian J. Braun, Andrés Carbonell Adames, Dieter Saur, Roland Rad
Rok vydání:	2022
Předmět:	Mice Phenotype Animals Clustered Regularly Interspaced Short Palindromic Repeats Genetic Testing CRISPR-Cas Systems General Biochemistry Genetics and Molecular Biology Gene Library
Zdroj:	Nature Protocols. 17:1903-1925
ISSN:	1750-2799 1754-2189
Popis:	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.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::de8119413100e5f3e5ec5c29ce0d7741 https://doi.org/10.1038/s41596-022-00700-y Zobrazit plný text záznamu Full text from SpringerLink