A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting

Kazuto Kobayashi; Tamae Ohye; Ira Pastan; Toshiharu Nagatsu

doi:10.1093/nar/24.18.3653

A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting

Kazuto Kobayashi, Tamae Ohye, Ira Pastan, Toshiharu Nagatsu

Regulatory Science

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

7 Citations (Scopus)

Abstract

Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor α subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation.

Original language	English
Pages (from-to)	3653-3655
Number of pages	3
Journal	Nucleic acids research
Volume	24
Issue number	18
DOIs	https://doi.org/10.1093/nar/24.18.3653
Publication status	Published - 1996

All Science Journal Classification (ASJC) codes

Genetics

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10.1093/nar/24.18.3653

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title = "A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting",

abstract = "Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor α subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation.",

author = "Kazuto Kobayashi and Tamae Ohye and Ira Pastan and Toshiharu Nagatsu",

note = "Funding Information: We thank T. Honjo for human IL-2Rα cDNA, T. Uchiyama for anti-Tac monoclonal antibody, and H. Sawada for technical instruction of immunofluorescence. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.",

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doi = "10.1093/nar/24.18.3653",

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T1 - A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting

AU - Kobayashi, Kazuto

AU - Ohye, Tamae

AU - Pastan, Ira

AU - Nagatsu, Toshiharu

N1 - Funding Information: We thank T. Honjo for human IL-2Rα cDNA, T. Uchiyama for anti-Tac monoclonal antibody, and H. Sawada for technical instruction of immunofluorescence. This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.

PY - 1996

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N2 - Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor α subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation.

AB - Immunotoxoin-mediated cell targeting (IMCT) is a technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor α subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation.

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A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting

Abstract

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