Efficient gene editing in induced pluripotent stem cells enabled by an inducible adenine base editor with tunable expression.
Title | Efficient gene editing in induced pluripotent stem cells enabled by an inducible adenine base editor with tunable expression. |
Publication Type | Journal Article |
Year of Publication | 2023 |
Authors | Nandy K, Babu D, Rani S, Joshi G, Ijee S, George A, Palani D, Premkumar C, Rajesh P, Vijayanand S, David E, Murugesan M, Velayudhan SR |
Journal | Sci Rep |
Volume | 13 |
Issue | 1 |
Pagination | 21953 |
Date Published | 2023 Dec 11 |
ISSN | 2045-2322 |
Keywords | Adenine, CRISPR-Cas Systems, Gene Editing, Induced Pluripotent Stem Cells, Mutation |
Abstract | The preferred method for disease modeling using induced pluripotent stem cells (iPSCs) is to generate isogenic cell lines by correcting or introducing pathogenic mutations. Base editing enables the precise installation of point mutations at specific genomic locations without the need for deleterious double-strand breaks used in the CRISPR-Cas9 gene editing methods. We created a bulk population of iPSCs that homogeneously express ABE8e adenine base editor enzyme under a doxycycline-inducible expression system at the AAVS1 safe harbor locus. These cells enabled fast, efficient and inducible gene editing at targeted genomic regions, eliminating the need for single-cell cloning and screening to identify those with homozygous mutations. We could achieve multiplex genomic editing by creating homozygous mutations in very high efficiencies at four independent genomic loci simultaneously in AAVS1-iABE8e iPSCs, which is highly challenging with previously described methods. The inducible ABE8e expression system allows editing of the genes of interest within a specific time window, enabling temporal control of gene editing to study the cell or lineage-specific functions of genes and their molecular pathways. In summary, the inducible ABE8e system provides a fast, efficient and versatile gene-editing tool for disease modeling and functional genomic studies. |
DOI | 10.1038/s41598-023-42174-2 |
Alternate Journal | Sci Rep |
PubMed ID | 38081875 |
PubMed Central ID | PMC10713686 |
Grant List | / WT_ / Wellcome Trust / United Kingdom IA/S/17/1/503118 / WTDBT_ / DBT-Wellcome Trust India Alliance / India |