Multiplexed Cre-dependent selection yields systemic AAVs for targeting distinct brain cell types.

Autor: Ravindra Kumar S; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Miles TF; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Chen X; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Brown D; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Dobreva T; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Huang Q; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.; Stanley Center for Psychiatric Research, Broad Institute, Cambridge, MA, USA., Ding X; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Luo Y; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Einarsson PH; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Greenbaum A; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.; Joint Department of Biomedical Engineering, North Carolina State University, Raleigh, NC, USA.; University of North Carolina at Chapel Hill, Chapel Hill, NC, USA., Jang MJ; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA., Deverman BE; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.; Stanley Center for Psychiatric Research, Broad Institute, Cambridge, MA, USA., Gradinaru V; Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA. viviana@caltech.edu.
Jazyk: angličtina
Zdroj: Nature methods [Nat Methods] 2020 May; Vol. 17 (5), pp. 541-550. Date of Electronic Publication: 2020 Apr 20.
DOI: 10.1038/s41592-020-0799-7
Abstrakt: Recombinant adeno-associated viruses (rAAVs) are efficient gene delivery vectors via intravenous delivery; however, natural serotypes display a finite set of tropisms. To expand their utility, we evolved AAV capsids to efficiently transduce specific cell types in adult mouse brains. Building upon our Cre-recombination-based AAV targeted evolution (CREATE) platform, we developed Multiplexed-CREATE (M-CREATE) to identify variants of interest in a given selection landscape through multiple positive and negative selection criteria. M-CREATE incorporates next-generation sequencing, synthetic library generation and a dedicated analysis pipeline. We have identified capsid variants that can transduce the central nervous system broadly, exhibit bias toward vascular cells and astrocytes, target neurons with greater specificity or cross the blood-brain barrier across diverse murine strains. Collectively, the M-CREATE methodology accelerates the discovery of capsids for use in neuroscience and gene-therapy applications.
Databáze: MEDLINE
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