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ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems
- Source :
- BMC Biology, BMC Biology, Vol 18, Iss 1, Pp 1-14 (2020)
- Publication Year :
- 2020
- Publisher :
- BioMed Central, 2020.
-
Abstract
- BackgroundMany favorable traits of crops and livestock and human genetic diseases arise from multiple single nucleotide polymorphisms or multiple point mutations with heterogeneous base substitutions at the same locus. Current cytosine or adenine base editors can only accomplish C-to-T (G-to-A) or A-to-G (T-to-C) substitutions in the windows of target genomic sites of organisms; therefore, there is a need to develop base editors that can simultaneously achieve C-to-T and A-to-G substitutions at the targeting site.ResultsIn this study, a novel fusion adenine and cytosine base editor (ACBE) was generated by fusing a heterodimer of TadA (ecTadAWT/*) and an activation-induced cytidine deaminase (AID) to the N- and C-terminals of Cas9 nickase (nCas9), respectively. ACBE could simultaneously induce C-to-T and A-to-G base editing at the same target site, which were verified in HEK293-EGFP reporter cell line and 45 endogenous gene loci of HEK293 cells. Moreover, the ACBE could accomplish simultaneous point mutations of C-to-T and A-to-G in primary somatic cells (mouse embryonic fibroblasts and porcine fetal fibroblasts) in an applicable efficiency. Furthermore, the spacer length of sgRNA and the length of linker could influence the dual base editing activity, which provided a direction to optimize the ACBE system.ConclusionThe newly developed ACBE would expand base editor toolkits and should promote the generation of animals and the gene therapy of genetic diseases with heterogeneous point mutations.
- Subjects :
- Physiology
Sus scrofa
Locus (genetics)
Plant Science
Computational biology
Biology
Mammalian systems
General Biochemistry, Genetics and Molecular Biology
Cytosine
Mice
chemistry.chemical_compound
Fetus
Structural Biology
Simultaneous C-to-T and A-to-G conversions
Animals
Humans
Point Mutation
lcsh:QH301-705.5
Ecology, Evolution, Behavior and Systematics
Subgenomic mRNA
Gene Editing
Cas9
Adenine
Point mutation
Methodology Article
HEK 293 cells
Cell Biology
Cytidine deaminase
Fibroblasts
Embryo, Mammalian
Adenine and cytosine base editor (ACBE)
HEK293 Cells
chemistry
lcsh:Biology (General)
General Agricultural and Biological Sciences
Linker
Developmental Biology
Biotechnology
Subjects
Details
- Language :
- English
- ISSN :
- 17417007
- Volume :
- 18
- Database :
- OpenAIRE
- Journal :
- BMC Biology
- Accession number :
- edsair.doi.dedup.....92f1d512a1ad72834502d79468dc23f9