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

Fingerprint

Ribonucleoproteins
Dependovirus
Zinc Fingers
Transgenes
Proteins
Research Personnel
Genome
Technology
Genes
Gene Editing

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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

Manufacturing and delivering genome-editing proteins. / Liu, Jia; Liang, Ya jun; Ren, Pei ling; Gaj, Thomas.

Methods in Molecular Biology. Humana Press Inc., 2018. p. 253-273 (Methods in Molecular Biology; Vol. 1867).

Research output: Chapter in Book/Report/Conference proceedingChapter

Liu, J, Liang, YJ, Ren, PL & Gaj, T 2018, Manufacturing and delivering genome-editing proteins. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1867, Humana Press Inc., pp. 253-273. https://doi.org/10.1007/978-1-4939-8799-3_19
Liu J, Liang YJ, Ren PL, Gaj T. Manufacturing and delivering genome-editing proteins. In Methods in Molecular Biology. Humana Press Inc. 2018. p. 253-273. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8799-3_19
Liu, Jia ; Liang, Ya jun ; Ren, Pei ling ; Gaj, Thomas. / Manufacturing and delivering genome-editing proteins. Methods in Molecular Biology. Humana Press Inc., 2018. pp. 253-273 (Methods in Molecular Biology).
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