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Specific gene transfer to neurons, endothelial cells and hematopoietic progenitors with lentiviral vectors

Abstract

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.

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Figure 1: Characterization of targeted lentiviral vectors.
Figure 2: Selective and efficient transduction of primary endothelial cells by CD105-LV.
Figure 3: Transduction of primary human HPCs via CD133.
Figure 4: Specific gene transfer to primary neurons using GluA2/4-LV.
Figure 5: In vivo application of GluA2/4-LV.

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Acknowledgements

This work was supported by grants from the Priority Programme “Mechanisms of gene vector entry and persistence” of the Deutsche Forschungsgemeinschaft to C.J.B. and K.C. and from the 7th European Community programme project Persisting Transgenesis (Persist) to C.J.B. U.C.M. was supported by grants of the Deutsche Forschungsgemeinschaft (MU 1457/8-1) and the “Nationales Genomforschungsnetz” (01GS08128). T.A. is supported by the graduate study program GK1172 Biologicals of the Goethe University Frankfurt AM.

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B.A., T.A. and S.K. designed and performed experiments and contributed to writing of the manuscript. J.H., A.C., J.B., I.C.S., R.C.M. and H.P. performed experiments. R.E.K., U.K., I.C.D.J. and K.K. contributed protocols and reagents. C.H. and H.M. supervised work. U.C.M. supervised work and contributed to writing of the manuscript. K.C. acquired grants. C.J.B. conceived and designed the study, acquired grants, supervised work and wrote the manuscript.

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Correspondence to Christian J Buchholz.

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Competing interests

I.C.D.J. is an employee of Miltenyi Biotec GmbH. S.K., K.C. and C.J.B. are listed as inventors in an international Patent Cooperation Treaty European patent application (PCT/EP2007/008384) assigned to the Paul Ehrlich Institut, which includes as claims the generation of targeted lentiviral vectors.

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Anliker, B., Abel, T., Kneissl, S. et al. Specific gene transfer to neurons, endothelial cells and hematopoietic progenitors with lentiviral vectors. Nat Methods 7, 929–935 (2010). https://doi.org/10.1038/nmeth.1514

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