pcmv pe2 plasmid expressing pe2 cas9 rt fusion protein  (Addgene inc)

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 iPE, iPEmax, nu-iPE, nu-iPEmax, twinPE, SpG-PE, SpRY-PE, nu-ciPE, ciPE, and Rep-X-assisted ciPE vectors were derived from the pCMV-PE2 vector (Addgene #132775) using a Uniclone One Step Seamless Cloning Kit (Genesand, CN).

Techniques: Variant Assay, Ligation, Reverse Transcription, Binding Assay, Comparison, Two Tailed Test

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 Rep-X-assisted ciPE. Rep-X is included to aid in unwinding DNA to enhance editing efficiency. b Schematic diagrams of structure of Rep-X-assisted ciPE editors. For abbreviations, see Fig. . c Comparison of inverse prime editing efficiencies between Rep-X-assisted ciPE2 and ciPE2 at six target sites in HEK293T cells. Frequencies (mean ± s.e.m.) at HEK4 , DMD , BCL11A , PSMB2, and GFAP sites were obtained from eight biological replicates ( n = 8) and three biological replicates ( n = 3) at HEXA site. d Comparison of inverse prime editing efficiencies between three ciPE constructs and three Rep-X-assisted ciPE constructs at the GFAP , HEK4 , and DMD target sites in HEK293T cells. Frequencies (mean ± s.e.m.) were obtained from eight biological replicates ( n = 8). e Inverse prime editing efficiencies at the DMD and GFAP sites in HEK293T cells using Rep-X-assisted ciPE4 and ciPE5 editors. Frequencies (mean ± s.e.m.) were obtained from three biological replicates ( n = 3). f Inverse prime editing efficiencies at the HEK4 and DMD sites in HeLa, K562, and U2OS cells using Rep-X-assisted ciPE4 and ciPE5 editors. Frequencies (mean ± s.e.m.) were obtained from four biological replicates ( n = 4). 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 iPE, iPEmax, nu-iPE, nu-iPEmax, twinPE, SpG-PE, SpRY-PE, nu-ciPE, ciPE, and Rep-X-assisted ciPE vectors were derived from the pCMV-PE2 vector (Addgene #132775) using a Uniclone One Step Seamless Cloning Kit (Genesand, CN).

Techniques: Comparison, Construct, Two Tailed Test