NON VIRAL GENE DELIVERY TO HEMATOPOIETIC STEM CELLS

非病毒基因递送至造血干细胞

• 批准号: 2417225
• 负责人: QUAN Karen ZHU
• 金额: $ 10万
• 依托单位: INTRAIMMUNE THERAPIES, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2417225
• 关键词: CD34 molecule; biotechnology; cell line; chimeric proteins; flow cytometry; gene targeting; green fluorescent proteins; hematopoietic stem cells; method development; nucleic acid chemical synthesis; peptide chemical synthesis; plasmids; protamines; protein purification; transfection; transfection /expression vector; CD34 molecule; biotechnology; cell line; chimeric proteins; flow cytometry; gene targeting; green fluorescent proteins; hematopoietic stem cells; method development; nucleic acid chemical synthesis; peptide chemical synthesis; plasmids; protamines; protein purification; transfection; transfection /expression vector

DESCRIPTION: (Adapted from the Applicant's Abstract). Hematopoietic stem cell are the ideal target for gene therapy of a variety of inherited and other disorders of the hematopoietic system. Administration of therapeutic genes to hematopoietic stem cells will provide a more robust treatment for disease than the introduction of genes into the progeny of cell types that are affected. To target gene delivery to hematopoietic stem cells, the investigators propose to develop a non-viral DNA delivery system capable of transferring DNA to human hematopoietic stem cells. The system uses a fusion protein that contains an antibody moiety directed against the hematopoietic stem cell specific surface marker CD34 and a DNA binding moiety. Four human anti-CD34 antibodies have been cloned as Fab genes, and will be used to synthesize fusion proteins with the human DNA binding protein protamine. This will be used to deliver plasmids containing marker genes to CD34-expressing cells. The ultimate goal is to develop a gene delivery vehicle that can be administered to patients in vivo. In this proposal, genes encoding fusion proteins will be synthesized, the protein will be expressed in sufficient quantities to test efficacy in cell culture, and gene delivery will be evaluated in cultured and primary CD34+ cells.

描述：（根据申请人的摘要进行了改编）。 造血茎 细胞是各种遗传和 造血系统的其他疾病。 管理 造血干细胞的治疗基因将提供更健壮的 疾病的治疗比将基因引入到后代的治疗 受影响的细胞类型。 靶向基因递送到造血 干细胞，研究人员建议开发非病毒DNA递送 能够将DNA转移到人造血干细胞的系统。 这 系统使用融合蛋白，该蛋白包含定向抗体部分的融合蛋白 针对造血细胞特异性表面标记CD34和A DNA结合部分。 四种人类抗CD34抗体被克隆为 Fab基因，将用于与人合成融合蛋白 DNA结合蛋白质蛋白质。 这将用于输送质粒 包含表达CD34的细胞的标记基因。 最终目标是 开发一个基因输送工具，可以对患者进行管理 体内。 在此建议中，编码融合蛋白的基因将是 合成的，该蛋白质将以足够的数量表示 在细胞培养中的测试功效和基因递送将在 培养和原代CD34+细胞。