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 language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Publisher | Humana Press Inc. |
| Pages | 253-273 |
| Number of pages | 21 |
| DOIs | |
| State | Published - Jan 1 2018 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1867 |
| 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