《Table 2–In vitro TLR-characterized targeting efficiency of U6 or U3 promoters as verified by in viv

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《Systematic identification of endogenous RNA polymerase Ⅲ promoters for efficient RNA guidebased genome editing technologies in maize》

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For three potential promoters（U6-2，U6-4，and U6-6）showing higher mutation frequencies，in vivo verification was produced for U6-2.More experiments should be performed to verify all the contrasting frequencies of U6 and U3 promoters from the different maize endogenous Pol III promoters observed in this study.In our previous study[14]，RNA-guided Cas9 based on the in vivo U6-6 promoter yielded efficiencies of up to 91.2%.We also obtained in vivo data for the important of Pol III promoters，U6-4，with efficiencies ranging from 42.9%to 78.9%on a RGEN-targeting Zm AGO1 locus（detailed data will be published in a separate paper），an important target gene for small RNA processing[16].Table 2 shows our in vitro data on mutation frequencies and summarizes in vivo mutation frequencies found using maize endogenous RNA Pol III promoters.The few reported results of in vivo gene targeting efficiency are consistent with our findings.Our data support the establishment of an efficient genome editing technology system relying on the RNA Pol III promoters for expressing single-guide RNAs.