A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting.
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 alpha 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.
Kobayashi
K
K
Institute for Comprehensive Medical Science, School of Medicine, Fujita Health University, Aichi, Japan.
Ohye
T
T
Pastan
I
I
Nagatsu
T
T
D036002:ADP Ribose Transferases; D000818:Animals; D001427:Bacterial Toxins; D002469:Cell Separation / Q000379:methods*; D005098:Exotoxins; D005240:Feasibility Studies; D005822:Genetic Vectors; D006801:Humans; D000922:Immunotoxins; D051379:Mice; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011549:Pseudomonas; D015375:Receptors, Interleukin-2 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D005098:Exotoxins; D000922:Immunotoxins; D015375:Receptors, Interleukin-2; D037521:Virulence Factors; D000097668:Pseudomonas aeruginosa Exotoxin A