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Cited by 9CRISPR and Genetic EngineeringRNA regulation and diseasePluripotent Stem Cells Research

PINE-TREE enables highly efficient genetic modification of human cell lines.

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Frisch, Carlye, Kostes, William W., Galyon, Brooke, Whitman, Brycelyn, Tekel, Stefan J., Standage-Beier, Kylie, Srinivasan, Gayathri, Wang, Xiao +1 more·2023·Molecular Therapy - Nucleic Acids
DOI 10.1016/j.omtn.2023.07.007PMC PMC10425837
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Abstract

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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Cited by

Sorted by citations

Authors
Frisch, C., Kostes, W. W., Galyon, B., Whitman, B., Tekel, S. J., Standage-Beier, K., Srinivasan, G., Wang, X., & Brafman, D. A.
Affiliations
Arizona State University (US)
Corresponding author
Brafman, David A.
ORCID
Venue
Molecular Therapy - Nucleic Acids
Publisher
Elsevier BV
Published
2023-09
Volume
33
Pages
483-492
ISSN
2162-2531 (ISSN-L)
DOI
10.1016/j.omtn.2023.07.007
PMC
PMC10425837
OpenAlex
W4384564858
Cited by
9
References
5
Citations by year
2026: 4 · 2025: 3 · 2024: 1 · 2023: 1
FWCI
0.78
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Therapy — Nucleic Acids (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Institutes of Health
  • National Institute on Aging (R21AG056706, R21AG075612, R21AG079279)
  • National Institute of General Medical Sciences (R01GM106081, R01GM121698)
  • National Institute of Biomedical Imaging and Bioengineering (R01EB031893)
  • Alzheimer's Association
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
prime editing, pine, genome editing efficiency, indels, human cell lines

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