Nature communications, 2018-08, Vol.9 (1), p.3048-10, Article 3048
2018
Volltextzugriff (PDF)
Details
- Autor(en) / Beteiligte
- Titel
- Directed evolution of CRISPR-Cas9 to increase its specificity
- Ist Teil von
- Nature communications, 2018-08, Vol.9 (1), p.3048-10, Article 3048
- Ort / Verlag
- England: Nature Publishing Group
- Erscheinungsjahr
- 2018
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- The use of CRISPR-Cas9 as a therapeutic reagent is hampered by its off-target effects. Although rationally designed S. pyogenes Cas9 (SpCas9) variants that display higher specificities than the wild-type SpCas9 protein are available, these attenuated Cas9 variants are often poorly efficient in human cells. Here, we develop a directed evolution approach in E. coli to obtain Sniper-Cas9, which shows high specificities without killing on-target activities in human cells. Unlike other engineered Cas9 variants, Sniper-Cas9 shows WT-level on-target activities with extended or truncated sgRNAs with further reduced off-target activities and works well in a preassembled ribonucleoprotein (RNP) format to allow DNA-free genome editing.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2041-1723eISSN: 2041-1723DOI: 10.1038/s41467-018-05477-xTitel-ID: cdi_doaj_primary_oai_doaj_org_article_12e41efab7774824b3a582ae5751b26d
- Format
- –
- Schlagworte
- CRISPR, CRISPR-Cas Systems, Deoxyribonucleic acid, Directed evolution, Directed Molecular Evolution, DNA, DNA - genetics, E coli, Escherichia coli - genetics, Evolution, Gene Editing, Genome editing, Genomes, HEK293 Cells, Humans, Plasmids - metabolism, Proteins, Reagents, Recombinant Proteins - chemistry, Ribonucleoproteins - chemistry, RNA, Guide, CRISPR-Cas Systems - genetics, Substrate Specificity