Examination of Factors Affecting Site-Directed RNA Editing by the MS2-ADAR1 Deaminase System
نویسندگان
چکیده
Adenosine deaminases acting on RNA (ADARs) have double-stranded binding domains and a deaminase domain (DD). We used the MS2 system specific guide RNAs to direct ADAR1-DD target adenosines in mRNA encoding-enhanced green fluorescence protein. Using this transfected HEK-293 cells, we evaluated effects of changing length position efficiency conversion amber (TAG) ochre (TAA) stop codons tryptophan (TGG) target. Guide 19, 21 23 nt were positioned upstream downstream MS2-RNA, providing total six RNAs. The more functionally effective than RNAs, with following hierarchy efficiency: > 19 nt. highest editing was 16.6%. Off-target not detected complementary region but 50 proportional amount inversely RNA. Our results suggest that requires precise optimization ratio enzyme, RNA,
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ژورنال
عنوان ژورنال: Genes
سال: 2023
ISSN: ['2073-4425']
DOI: https://doi.org/10.3390/genes14081584