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

Carlye Frisch; William W. Kostes; Brooke Galyon; Brycelyn Whitman; Stefan J. Tekel; Kylie Standage-Beier; Gayathri Srinivasan; Xiao Wang; David A. Brafman

doi:10.1016/j.omtn.2023.07.007

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

Carlye Frisch, William W. Kostes, Brooke Galyon, Brycelyn Whitman, Stefan J. Tekel, Kylie Standage-Beier, Gayathri Srinivasan, Xiao Wang, David A. Brafman

Engineering, Ira A. Fulton Schools of (IAFSE)

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

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.

Original language	English (US)
Pages (from-to)	483-492
Number of pages	10
Journal	Molecular Therapy Nucleic Acids
Volume	33
DOIs	https://doi.org/10.1016/j.omtn.2023.07.007
State	Published - Sep 12 2023

Keywords

CRISPR
MT: RNA/DNA
genome modification
human pluripotent stem cells
prime editing

ASJC Scopus subject areas

Molecular Medicine
Drug Discovery

Access to Document

10.1016/j.omtn.2023.07.007

Cite this

@article{0ae3e2ef12f44a798df36d08e3259ef9,

title = "PINE-TREE enables highly efficient genetic modification of human cell lines",

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.",

keywords = "CRISPR, MT: RNA/DNA, genome modification, human pluripotent stem cells, prime editing",

author = "Carlye Frisch and Kostes, {William W.} and Brooke Galyon and Brycelyn Whitman and Tekel, {Stefan J.} and Kylie Standage-Beier and Gayathri Srinivasan and Xiao Wang and Brafman, {David A.}",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors",

year = "2023",

month = sep,

day = "12",

doi = "10.1016/j.omtn.2023.07.007",

language = "English (US)",

volume = "33",

pages = "483--492",

journal = "Molecular Therapy Nucleic Acids",

issn = "2162-2531",

publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - PINE-TREE enables highly efficient genetic modification of human cell lines

AU - Frisch, Carlye

AU - Kostes, William W.

AU - Galyon, Brooke

AU - Whitman, Brycelyn

AU - Tekel, Stefan J.

AU - Standage-Beier, Kylie

AU - Srinivasan, Gayathri

AU - Wang, Xiao

AU - Brafman, David A.

PY - 2023/9/12

Y1 - 2023/9/12

N2 - 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.

AB - 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.

KW - CRISPR

KW - MT: RNA/DNA

KW - genome modification

KW - human pluripotent stem cells

KW - prime editing

UR - http://www.scopus.com/inward/record.url?scp=85169031403&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85169031403&partnerID=8YFLogxK

U2 - 10.1016/j.omtn.2023.07.007

DO - 10.1016/j.omtn.2023.07.007

M3 - Article

AN - SCOPUS:85169031403

SN - 2162-2531

VL - 33

SP - 483

EP - 492

JO - Molecular Therapy Nucleic Acids

JF - Molecular Therapy Nucleic Acids

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this