Fig. 2
From: Efficient and versatile multiplex prime editing in hexaploid wheat
Engineered prime editors by mutating RT and changing the prime editor protein architecture. a Schematic representation of M-MLV RT without the RNase H domain structure showing the functionally important amino acids residues including F156 in the palm region, V223 in the palm region and F309 in the thumb region that we mutated. b Frequencies of prime editing and byproducts induced by ePPE, ePPE-F156W, ePPE-V223A, ePPE-V223H, ePPE-V223I, ePPE-F309N, and ePPE-V223H-F309N prime editors with epegRNAs. c Summary of the fold change in prime editing efficiencies for engineered ePPEs compared to ePPE. Values were calculated from the data presented in b. The editing frequencies using ePPE for each target were normalized to 1, and the frequencies using other ePPEs for each target were adjusted accordingly. d Schematic representation of ePPE, ePPE*, ePPEmax, and ePPEmax*. NLSSV40, the SV40 NLS; NLSc−Myc, the c-Myc NLS; bpNLSSV40, the bipartite SV40 NLS; vbpNLSSV40, a variant of bipartite SV40 NLS. e Frequencies of prime editing and byproducts induced by ePPE, ePPE*, ePPEmax, and ePPEmax*. f Summary of the fold change in prime editing efficiencies for ePPE*, ePPEmax and ePPEmax* compared to ePPE. Values were calculated from the data presented in e. The editing frequencies using ePPE for each target were normalized to 1, and the frequencies using other ePPEs for each target were adjusted accordingly. Frequencies (mean ± s.e.m.) were calculated from three independent experiments (n = 3) in b and e. P values were obtained using the two-tailed Student’s t test in c and f: *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001