Journal: Nature Communications
Article Title: Circular RNA-mediated inverse prime editing in human cells
doi: 10.1038/s41467-025-59120-7
Figure Lengend Snippet: a Schematic diagram of circular RNA-mediated inverse prime editors (ciPEs). b Schematic diagrams of the structure of ciPE editors. CMV, the CMV promoter of cytomegalovirus; U6, the polymerase III promoter of U6; NLS, bipartite nuclear localization signal; M-MLV RTΔRNase H, deletion variant of M-MLV RT with no RNase H domain; 5′ RL, 5′ ribozyme and ligation sequences; 3′ RL, 3′ ribozyme and ligation sequences; RTT, reverse transcriptase template; PBS, primer binding site. c Comparison of inverse prime editing frequencies between split iPE2 and ciPE2 at four target sites in HEK293T cells. Frequencies (mean ± s.e.m.) in c were obtained from three biological replicates ( n = 3). d Comparison of inverse prime editing efficiencies between ciPE2–5 and split iPEmax2–5 using epegRNA at six target sites in HEK293T cells. Frequencies (mean ± s.e.m.) in d were obtained from four biological replicates ( n = 4). circRNA, circular RNA; Ins, insertion; Del, deletion. InDels, byproducts of random insertions and deletions. P values were obtained from two-tailed Student’s t -test: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Source data are provided as a Source Data file.
Article Snippet: The sgRNA, nicking sgRNA, pegRNA, and epegRNA vectors driven by the human U6 promoter were constructed by annealing oligonucleotides and inserting them into the pOsU3 backbone (Addgene #170132) using the Golden Gate assembly method.
Techniques: Variant Assay, Ligation, Reverse Transcription, Binding Assay, Comparison, Two Tailed Test
Addgene inc is a verified supplier 
