CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells
| العنوان: | CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells |
|---|---|
| المؤلفون: | J. Keith Joung, Ronghao Zhou, Sowmya Iyer, Bret R. Miller, Lukas M. Langner, Ibrahim C. Kurt, Julian Grünewald, Sara P. Garcia |
| المصدر: | Nature biotechnology |
| بيانات النشر: | Springer Science and Business Media LLC, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0303 health sciences, Cas9, APOBEC1, Biomedical Engineering, RNA, Bioengineering, Computational biology, Cytidine deaminase, Applied Microbiology and Biotechnology, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, chemistry, Molecular Medicine, CRISPR, Indel, 030217 neurology & neurosurgery, Cytosine, DNA, 030304 developmental biology, Biotechnology |
| الوصف: | CRISPR-guided DNA cytosine and adenine base editors (CBEs and ABEs) are widely used for many applications1–4 but primarily create DNA base transitions (i.e., pyrimidine-to-pyrimidine, or purine-to-purine). Here we describe the engineering of two base editor architectures that can efficiently induce targeted C-to-G base transversions, with reduced levels of unwanted C-to-W (W = A or T) and indel mutations. One of these C-to-G base editors (CGBE1), consists of an RNA-guided Cas9 nickase, an E. coli-derived uracil DNA N-glycosylase (eUNG), and a rat APOBEC1 cytidine deaminase variant (R33A) previously shown to have reduced off-target RNA and DNA editing activities5, 6. We show that CGBE1 can efficiently induce C-to-G edits, particularly in AT-rich sequence contexts in human cells. We also removed the eUNG domain to yield miniCGBE1, which reduced indel frequencies but only modestly decreased editing efficiency. CGBE1 and miniCGBE1 enable C-to-G edits and will serve as a basis for optimizing C-to-G base editors for research and therapeutic applications. Editorial summary A new base editor enables the creation of C-to-G base changes in human cells. |
| تدمد: | 1546-1696 1087-0156 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c55363855f17e643e094eef4db9a52e3 https://doi.org/10.1038/s41587-020-0609-x |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....c55363855f17e643e094eef4db9a52e3 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 15461696 10870156 |
|---|