Application of CRISPR/Cas9 Pooled Screening for a Non-immediate Readout Model


TERZİ ÇİZMECİOĞLU N.

Methods in molecular biology (Clifton, N.J.), vol.2849, pp.87-116, 2024 (Scopus) identifier identifier

  • Publication Type: Article / Article
  • Volume: 2849
  • Publication Date: 2024
  • Doi Number: 10.1007/7651_2024_529
  • Journal Name: Methods in molecular biology (Clifton, N.J.)
  • Journal Indexes: Scopus, Biotechnology Research Abstracts, CAB Abstracts, MEDLINE
  • Page Numbers: pp.87-116
  • Keywords: Cas9, CRISPR, Differentiation, Neuroectoderm, Pooled screening, Sox1GFP
  • Middle East Technical University Affiliated: Yes

Abstract

Linking phenotypes to genetic components has been an essential part of novel drug discovery, and screening methods have been widely employed to achieve such a goal. Screens can be conducted in either pooled or arrayed formats. Although arrayed screenings provide a better and cheaper alternative in small scale, the larger-scale screenings are conducted in pooled manner. With its adaptability to various models and conditions, CRISPR/Cas9 technology provides an invaluable alternative to classical and RNAi-based screening methods. Combined with high-throughput sequencing and bioinformatics, CRISPR-/Cas9-based pooled screening methods provide unbiased and robust data. In this protocol, we employed CRISPR-/Cas9-based pooled screening for a non-binary and non-immediate readout.