Surrogate reporters for enrichment of cells with nuclease-induced mutations

Hyojin Kim; Eunji Um; Sung Rae Cho; Chorong Jung; Hyongbum Kim; Jin Soo Kim

doi:10.1038/nmeth.1733

Surrogate reporters for enrichment of cells with nuclease-induced mutations

Hyojin Kim, Eunji Um, Sung Rae Cho, Chorong Jung, Hyongbum Kim, Jin Soo Kim

Department of Rehabilitation Medicine

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

166 Citations (Scopus)

Abstract

Zinc-finger nucleases (ZFNs) and TAL-effector nucleases (TALENs) are powerful tools for creating genetic modifications in eukaryotic cells and organisms. But wild-type and mutant cells that contain genetic modifications induced by these programmable nucleases are often phenotypically indistinguishable, hampering isolation of mutant cells. Here we show that transiently transfected episomal reporters encoding fluorescent proteins can be used as surrogate genes for the efficient enrichment of endogenous gene-modified cells by flow cytometry.

Original language	English
Pages (from-to)	941-943
Number of pages	3
Journal	Nature Methods
Volume	8
Issue number	11
DOIs	https://doi.org/10.1038/nmeth.1733
Publication status	Published - 2011 Nov

Bibliographical note

Funding Information:
J.-S.K. and Hyongbum K. are supported in part by the National Research Foundation of Korea (J.-S.K., 2011-0000402; Hyongbum K., 2011-0013568, 2011-27177, 2011-0019357 and 2011-0028267).

All Science Journal Classification (ASJC) codes

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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10.1038/nmeth.1733

Cite this

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abstract = "Zinc-finger nucleases (ZFNs) and TAL-effector nucleases (TALENs) are powerful tools for creating genetic modifications in eukaryotic cells and organisms. But wild-type and mutant cells that contain genetic modifications induced by these programmable nucleases are often phenotypically indistinguishable, hampering isolation of mutant cells. Here we show that transiently transfected episomal reporters encoding fluorescent proteins can be used as surrogate genes for the efficient enrichment of endogenous gene-modified cells by flow cytometry.",

author = "Hyojin Kim and Eunji Um and Cho, {Sung Rae} and Chorong Jung and Hyongbum Kim and Kim, {Jin Soo}",

note = "Funding Information: J.-S.K. and Hyongbum K. are supported in part by the National Research Foundation of Korea (J.-S.K., 2011-0000402; Hyongbum K., 2011-0013568, 2011-27177, 2011-0019357 and 2011-0028267).",

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AU - Kim, Hyojin

AU - Um, Eunji

AU - Cho, Sung Rae

AU - Jung, Chorong

AU - Kim, Hyongbum

AU - Kim, Jin Soo

N1 - Funding Information: J.-S.K. and Hyongbum K. are supported in part by the National Research Foundation of Korea (J.-S.K., 2011-0000402; Hyongbum K., 2011-0013568, 2011-27177, 2011-0019357 and 2011-0028267).

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AB - Zinc-finger nucleases (ZFNs) and TAL-effector nucleases (TALENs) are powerful tools for creating genetic modifications in eukaryotic cells and organisms. But wild-type and mutant cells that contain genetic modifications induced by these programmable nucleases are often phenotypically indistinguishable, hampering isolation of mutant cells. Here we show that transiently transfected episomal reporters encoding fluorescent proteins can be used as surrogate genes for the efficient enrichment of endogenous gene-modified cells by flow cytometry.

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Surrogate reporters for enrichment of cells with nuclease-induced mutations

Abstract

Bibliographical note

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