TitleProtocol for arrayed gRNA screening by base editors in mammalian cell lines using lentiviral system.
Publication TypeJournal Article
Year of Publication2023
AuthorsRavi NSam, George A, Mohankumar KM
JournalSTAR Protoc
Date Published2023 Dec 15
KeywordsAnimals, Cell Line, CRISPR-Cas Systems, Gene Editing, Lentivirus, Mammals, RNA, Guide, CRISPR-Cas Systems

Base editing, a CRISPR-based genome engineering technique, enables precise single-nucleotide modifications while minimizing double-strand breaks. Here, we present a protocol for arrayed mutagenesis using base editors to identify regulatory elements within the gamma-globin locus. We describe steps for guide RNA (gRNA) cloning into lentiviral vectors, establishing stable cell lines with base editor expression, transducing gRNAs, and assessing editing efficiency. This protocol can be applied to diverse genomic regions and cell lines for arrayed screening, facilitating genetic research, and target discovery. For complete details on the use and execution of this protocol, please refer to Ravi et al. (2022).

Alternate JournalSTAR Protoc
PubMed ID37922314
PubMed Central IDPMC10656259