Journal: Nucleic Acids Research
Article Title: Triple base editor catalyzes saturation mutation of adenine, cytidine, and guanine
doi: 10.1093/nar/gkaf1423
Figure Lengend Snippet: Design and screening of triple base editors (ACG-BEs). ( a ) The process of ACG-BEs catalyzes three types of base substrates (A, C, and G) to generate DNA sequence diversity. Adenosine deaminase refers to evolved Escherichia coli adenosine deaminase mutants (TadA-8e, T AD AC-3.1, T AD AC-3.155, and TadA-dual); nCas9, Cas9 D10A; MPG v6.3, derived from evolved N-methylpurine DNA glycosylase. ( b ) Schematics of the constructs for ACG-BEs. bNLS, bipartite nuclear localization signals; T AD AC-3.1, T AD AC-3.155, and TadA-dual, derived from evolved E. coli adenosine deaminase; nCas9, Cas9 D10A; MPG v3, and MPG v6.3, derived from evolved N-methylpurine DNA glycosylase; linkers are also shown. ( c ) Base-editing outcomes of ACG-BEs at the endogenous target PLS3-AS1 -sg1 in HEK293T cells. Data are means ± SD ( n = 3 independent experiments). ( d ) Allele table for PLS3-AS1 -sg1 in HEK293T cells after ACG-BE7 transfection. The target site allele is boxed in a gray line. The percentile and sequencing read of each allele at one representative of three independent experiments are listed on the right. ( e ) The allele base editing efficiency of ACG-BEs across the protospacer at the endogenous target PLS3-AS1- sg1 in HEK293T cells. Data are means ± SD ( n = 3 independent experiments).
Article Snippet: Human codon-optimized TadA-dual, T AD A-3.1, T AD A-3.155, MPGv3, and MPG v6.3 were synthesized by Genewiz (Suzhou, China).
Techniques: Sequencing, Derivative Assay, Construct, Transfection
