A G-CSF receptor-gyrase B fusion gene: A new type of molecular switch for expansion of genetically modified hematopoietic cells.
We have developed a novel system for expansion of transduced hematopoietic cells. This system involves "selective amplifier genes" encoding fusion proteins between the granulocyte colony-stimulating factor receptor (Gcr) and the estrogen receptor (Er). The GcrEr chimeric gene conferred estrogen-dependent growth ability on murine hematopoietic cells. Here, we constructed a modified "selective amplifier gene" to circumvent possible concerns with the Er/estrogen switching system. The bacterial gyrase B (Gyr) gene was fused to the Gcr gene, and the GcrGyr fusion construct was introduced into interleukin-3 (IL-3)-dependent Ba/F3 cells. The dimeric antibiotic coumermycin induced IL-3-independent growth in Ba/F3 cells expressing GcrGyr. This stimulatory effect was antagonized by an excess amount of novobiocin, a monomeric form of coumermycin. These results suggest the feasibility of using Gyr as a molecular switch to regulate a growth signal in hematopoietic cells.
Kume
A
A
Division of Genetic Therapeutics, Center for Molecular Medicine, Tochigi, 3311-1 Yakushiji, 329-0498, Japan.
Ito
K
K
Ueda
Y
Y
Hasegawa
M
M
Urabe
M
M
Mano
H
H
Ozawa
K
K
D049933:Aminocoumarins; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019677:Artificial Gene Fusion; D002452:Cell Count; D018929:Cell Culture Techniques; D002460:Cell Line; D002678:Chimera; D003374:Coumarins / Q000494:pharmacology; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D005786:Gene Expression Regulation; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D051379:Mice; D009675:Novobiocin / Q000494:pharmacology; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000235:genetics*; D013997:Time Factors; D014162:Transfection
D049933:Aminocoumarins; D000900:Anti-Bacterial Agents; D003374:Coumarins; D016188:Receptors, Granulocyte Colony-Stimulating Factor; D009675:Novobiocin; D027081:DNA Gyrase; D004250:DNA Topoisomerases, Type II; C004628:coumermycin