Manufacturing and delivering genome-editing proteins

Jia Liu, Ya jun Liang, Pei ling Ren, Thomas Gaj

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages253-273
Number of pages21
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1867
ISSN (Print)1064-3745

Keywords

  • CRISPR
  • Genome editing
  • Protein delivery
  • RNP
  • TALENs
  • ZFNs

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Liu, J., Liang, Y. J., Ren, P. L., & Gaj, T. (2018). Manufacturing and delivering genome-editing proteins. In Methods in Molecular Biology (pp. 253-273). (Methods in Molecular Biology; Vol. 1867). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8799-3_19