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Manufacturing and Delivering Genome-Editing Proteins

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Zinc Finger Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1867))

Abstract

Genome-editing technologies have revolutionized the biomedical sciences by providing researchers with the ability to quickly and efficiently modify genes. While programmable nucleases can be introduced into cells using a variety of techniques, their delivery as purified proteins is an effective approach for limiting off-target effects. Here, we describe step-by-step procedures for manufacturing and delivering genome-modifying proteins—including Cas9 ribonucleoproteins (RNPs) and TALE and zinc-finger nucleases—into mammalian cells. Protocols for combining Cas9 RNP with naturally recombinogenic adeno-associated virus (AAV) donor vectors for the seamless insertion of transgenes by homology-directed genome editing are also provided.

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Acknowledgments

This work was supported by Natural Science Foundation of China (No. 31600686 to J.L.) and ShanghaiTech University (Startup fund to the Laboratory of ADC Chemistry).

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Authors and Affiliations

  1. Shanghai Institute for Advanced Immunochemical Studies (SIAIS), ShanghaiTech University, Shanghai, China

    Jia Liu & Ya-jun Liang

  2. Shanghai Advanced Research Institute, Chinese Academy of Science, Shanghai, China

    Pei-ling Ren

  3. University of Chinese Academy of Sciences, Beijing, China

    Pei-ling Ren

  4. Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA

    Thomas Gaj

Authors
  1. Jia Liu
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  2. Ya-jun Liang
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  3. Pei-ling Ren
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  4. Thomas Gaj
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Corresponding authors

Correspondence to Jia Liu or Thomas Gaj .

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Editors and Affiliations

  1. Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai, China

    Jia Liu

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Liu, J., Liang, Yj., Ren, Pl., Gaj, T. (2018). Manufacturing and Delivering Genome-Editing Proteins. In: Liu, J. (eds) Zinc Finger Proteins. Methods in Molecular Biology, vol 1867. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8799-3_19

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  • DOI: https://doi.org/10.1007/978-1-4939-8799-3_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8798-6

  • Online ISBN: 978-1-4939-8799-3

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