RRC ID 21519
Author Kaneko E, Kawahara M, Ueda H, Nagamune T.
Title Growth control of genetically modified cells using an antibody/c-Kit chimera.
Journal J Biosci Bioeng
Abstract Gene therapy has been regarded as an innovative potential treatment against serious congenital diseases. However, applications of gene therapy remain limited, partly because its clinical success depends on therapeutic gene-transduced cells acquiring a proliferative advantage. To address this problem, we have developed the antigen-mediated genetically modified cell amplification (AMEGA) system, which uses chimeric receptors to enable the selective proliferation of gene-transduced cells. In this report, we describe mimicry of c-Kit signaling and its application to the AMEGA system. We created an antibody/c-Kit chimera in which the extracellular domain of c-Kit is replaced with an anti-fluorescein single-chain Fv antibody fragment and the extracellular D2 domain of the erythropoietin receptor. A genetically modified mouse pro-B cell line carrying this chimera showed selective expansion in the presence of fluorescein-conjugated BSA (BSA-FL) as a growth inducer. By further engineering the transmembrane domain of the chimera to reduce interchain interaction we attained stricter ligand-dependency. Since c-Kit is an important molecule in the expansion of hematopoietic stem cells (HSCs), this antibody/c-Kit chimera could be a promising tool for gene therapy targeting HSCs.
Volume 113(5)
Pages 641-6
Published 2012-5-1
DOI 10.1016/j.jbiosc.2011.12.005
PII S1389-1723(11)00520-2
PMID 22227120
MeSH Animals Antibodies / metabolism* Antigens / metabolism Cell Line Cell Proliferation Chimera / metabolism* Genetic Therapy Hematopoietic Stem Cells / cytology* Humans Mice Precursor Cells, B-Lymphoid Receptors, Erythropoietin / genetics Receptors, Erythropoietin / metabolism Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / immunology Recombinant Fusion Proteins / metabolism Signal Transduction Single-Chain Antibodies / genetics
IF 2.366
Times Cited 17
DNA material pUC kit EH1 (RDB01344)