TL;DR
A detailed protocol for performing gene editing using an adenine base editor (ABE) to induce a single A-G transition mutation in a gene of therapeutic relevance is provided, enabling the production of cell models suited to the study of both dominant and recessive mutations.
This chapter provides a practical protocol for base editing by electroporation by electroporation in adherent and suspension stem cells and details essential steps for cell handling and recovery, followed by standard readouts.
An improved method for efficient generation of multi-KOs using sgRNA DNA strands and the CRISPR/Cas9 system is presented, which offers a simpler, faster, and cost-effective approach for generating multigene knockout clones, with implications for functional genomics, therapeutic development, and synthetic biology.
DNA repair outcomes at nicks and mismatches introduced by base editors at the PCSK9 and HBG loci in human embryos are evaluated, finding that unlike Cas9-induced DNA breaks, base editor-induced lesions are efficiently repaired.
Background/Objectives: CRISPR/Cas-dependent base editing enables the precise correction of genomic sequences by direct modification of DNA bases without creating potentially mutagenic DNA double-strand breaks. Base editing is considered a therapeutic approach for rare monogenic disorders or viral infections. However, m...
This work provides the first functional analysis of double-strand break repair in a chelicerate of T. urticae, and demonstrates that Polθ-deficient strains enhance incorporation of repair templates, even when mutations are distant from the cut site.
A systematic, comprehensive and efficient workflow combining generation, genotyping and expansion of high-quality monoclonal iPSC lines following CRISPR-Cas9 genome editing, which provides a robust and high-throughput framework for the reliable production of monoclonal genome-edited iPSCs, thereby advancing translation...
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