AI Article Synopsis
- Base editing is a CRISPR technique that allows precise changes to single nucleotides with reduced DNA damage.
- The protocol outlines how to use base editors for mutagenesis to study regulatory elements in the gamma-globin locus.
- Steps include cloning gRNAs, creating stable cell lines, transducing gRNAs, and measuring editing effectiveness, suitable for various genomic regions and cell lines.
Article Abstract
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).
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10656259 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2023.102668 | DOI Listing |
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