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PINE-TREE enables highly efficient genetic modification of human cell lines.
Frisch, Carlye; Kostes, William W; Galyon, Brooke; Whitman, Brycelyn; Tekel, Stefan J; Standage-Beier, Kylie; Srinivasan, Gayathri; Wang, Xiao; Brafman, David A.
Afiliação
  • Frisch C; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Kostes WW; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Galyon B; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Whitman B; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Tekel SJ; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Standage-Beier K; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Srinivasan G; Molecular and Cellular Biology Graduate Program, Arizona State University, Tempe, AZ 85287, USA.
  • Wang X; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
  • Brafman DA; School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA.
Mol Ther Nucleic Acids ; 33: 483-492, 2023 Sep 12.
Article em En | MEDLINE | ID: mdl-37588683
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Mol Ther Nucleic Acids Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Mol Ther Nucleic Acids Ano de publicação: 2023 Tipo de documento: Article