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Light-responsive charge-reversal nanovector for high-efficiency in vivo CRISPR/Cas9 gene editing with controllable location and time

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Abstract

Controllably and efficaciously localized CRISPR/Cas9 plasmids transfection plays an essential role in genetic editing associated with various key human diseases. We employed near-infrared (NIR) light-responsive CRISPR/Cas9 plasmids delivery via a charge-reversal nanovector to achieve highly efficient and site-specific gene editing. The nanovector with abundant positive charges was fabricated on the basis of an ultraviolet-sensitive conjugated polyelectrolyte coated on an upconversion nanomaterial (UCNP-UVP-P), which can convert into negative charges upon 980 nm light irradiation. Using the as-prepared nanovector, we demonstrated the plasmids could be efficiently transfected into tumor cells (∼ 63% ± 4%) in a time-controlled manner, and that functional CRISPR/Cas9 proteins could be successfully expressed in a selected NIR-irradiated region. Particularly, this strategy was successfully applied to the delivery of CRISPR/Cas9 gene to tumor cells in vivo, inducing high efficiency editing of the target gene PLK-1 under photoirradiation. Therefore, this precisely controlled gene regulation strategy has the potential to serve as a new paradigm for gene engineering in complex biological systems.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (Nos. 21771065 and 81630046); the Guangdong Special Support Program (No. 2017TQ04R138), the Natural Science Foundation of Guangdong (No. 2019A1515012021), the Science and Technology Planning Project of Guangdong (No. 2017A020215088), Pearl River Nova Program of Guangzhou (No. 201806010189).

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  1. MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University, Guangzhou, 510631, China

    Yunxia Wu, Judun Zheng, Qin Zeng, Tao Zhang & Da Xing

  2. Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, 510631, China

    Yunxia Wu, Judun Zheng, Qin Zeng, Tao Zhang & Da Xing

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  1. Yunxia Wu
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  4. Tao Zhang
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Correspondence to Tao Zhang or Da Xing.

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12274_2020_2864_MOESM1_ESM.pdf

Light-responsive charge-reversal nanovector for high-efficiency in vivo CRISPR/Cas9 gene editing with controllable location and time

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Wu, Y., Zheng, J., Zeng, Q. et al. Light-responsive charge-reversal nanovector for high-efficiency in vivo CRISPR/Cas9 gene editing with controllable location and time. Nano Res. 13, 2399–2406 (2020). https://doi.org/10.1007/s12274-020-2864-z

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