| Autor: |
Carlye Frisch, William W. Kostes, Brooke Galyon, Brycelyn Whitman, Stefan J. Tekel, Kylie Standage-Beier, Gayathri Srinivasan, Xiao Wang, David A. Brafman |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
|
| Zdroj: |
Molecular Therapy: Nucleic Acids, Vol 33, Iss , Pp 483-492 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
2162-2531 |
| DOI: |
10.1016/j.omtn.2023.07.007 |
| Popis: |
Prime editing technologies enable precise genome editing without the caveats of CRISPR nuclease-based methods. Nonetheless, current approaches to identify and isolate prime-edited cell populations are inefficient. Here, we established a fluorescence-based system, prime-induced nucleotide engineering using a transient reporter for editing enrichment (PINE-TREE), for real-time enrichment of prime-edited cell populations. We demonstrated the broad utility of PINE-TREE for highly efficient introduction of substitutions, insertions, and deletions at various genomic loci. Finally, we employ PINE-TREE to rapidly and efficiently generate clonal isogenic human pluripotent stem cell lines, a cell type recalcitrant to genome editing. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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