Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
Nat Methods. 2010 Nov;7(11):929-35. doi: 10.1038/nmeth.1514. Epub 2010 Oct 10.
We present a flexible and highly specific targeting method for lentiviral vectors based on single-chain antibodies recognizing cell-surface antigens. We generated lentiviral vectors specific for human CD105(+) endothelial cells, human CD133(+) hematopoietic progenitors and mouse GluA-expressing neurons. Lentiviral vectors specific for CD105 or for CD20 transduced their target cells as efficiently as VSV-G pseudotyped vectors but discriminated between endothelial cells and lymphocytes in mixed cultures. CD133-targeted vectors transduced CD133(+) cultured hematopoietic progenitor cells more efficiently than VSV-G pseudotyped vectors, resulting in stable long-term transduction. Lentiviral vectors targeted to the glutamate receptor subunits GluA2 and GluA4 exhibited more than 94% specificity for neurons in cerebellar cultures and when injected into the adult mouse brain. We observed neuron-specific gene modification upon transfer of the Cre recombinase gene into the hippocampus of reporter mice. This approach allowed targeted gene transfer to many cell types of interest with an unprecedented degree of specificity.
我们提出了一种基于识别细胞表面抗原的单链抗体的灵活且高度特异的慢病毒载体靶向方法。我们生成了针对人 CD105(+)内皮细胞、人 CD133(+)造血祖细胞和表达 GluA 的小鼠神经元的慢病毒载体。针对 CD105 或 CD20 的慢病毒载体能够有效地转导其靶细胞，与 VSV-G 假型载体一样，但能够区分混合培养物中的内皮细胞和淋巴细胞。CD133 靶向载体比 VSV-G 假型载体更有效地转导 CD133(+)培养的造血祖细胞，从而实现稳定的长期转导。靶向谷氨酸受体亚基 GluA2 和 GluA4 的慢病毒载体对小脑培养物中的神经元具有超过 94%的特异性，并且当注入成年小鼠大脑时也是如此。我们观察到在报告小鼠的海马体中转移 Cre 重组酶基因后，神经元具有特异性基因修饰。这种方法允许以空前的特异性将靶基因转移到许多感兴趣的细胞类型。
