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 |
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Keywords
- CRISPR
- Genome editing
- Protein delivery
- RNP
- TALENs
- ZFNs
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
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 proceeding › Chapter
}
TY - CHAP
T1 - Manufacturing and delivering genome-editing proteins
AU - Liu, Jia
AU - Liang, Ya jun
AU - Ren, Pei ling
AU - Gaj, Thomas
PY - 2018/1/1
Y1 - 2018/1/1
N2 - 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.
AB - 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.
KW - CRISPR
KW - Genome editing
KW - Protein delivery
KW - RNP
KW - TALENs
KW - ZFNs
UR - http://www.scopus.com/inward/record.url?scp=85052672152&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85052672152&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-8799-3_19
DO - 10.1007/978-1-4939-8799-3_19
M3 - Chapter
C2 - 30155829
AN - SCOPUS:85052672152
T3 - Methods in Molecular Biology
SP - 253
EP - 273
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -