elisa assay  (R&D Systems)

Journal: Nature Communications

Article Title: Effective in vivo RNA base editing via engineered cytidine deaminase APOBECs fused with PUF proteins

doi: 10.1038/s41467-025-64748-6

Figure Lengend Snippet: a Determination of editing windows for CU5.1 on C 459 of EGFP mRNA. The graph shows on-target editing rates by CU5.1 at various distances from the ePUF10 binding site. b Editing efficacy of CU5.1 on C 526 of APOE (APOE4 & APOE3) reporter mRNA expressed in HEK293T cells. The ePUF10-binding sites are marked in blue. c Editing efficacy of CU5.1(Top) and the Cas13-bases xCBE (bottom) editors on C 526 of endogenous APOE3 mRNA in HepG2 cells. The ePUF10-binding site and gRNA-xCBE target regions are underlined. Editing efficacy of CU5.1 on C 143 of SOD1 reporter mRNA ( d ) and C 76 of Rhodopsin reporter mRNA ( e ) expressed in HEK293T cells. The ePUF10-binding sites are marked in blue. f Summary of the editing window for CU5.1 across EGFP, APOE4/3, SOD1 and Rhodopsin reporter mRNAs. The x -axis represents the distance from the editing site to the ePUF10 binding site; the y -axis shows the C-to-U editing rate. Editing rates were acquired by Sanger sequencing. g Determination of editing windows for CU5.15 on C 459 of EGFP mRNA. Editing efficacy of CU5.15 on C 104 of Mef2c reporter mRNA ( h ) and C 823 of PCSK9 reporter mRNA ( i ) expressed in HEK293T cells. j Summary of the cytosine editing window mediated by CU5.15 around the target sites of EGFP, PCSK9, and Mef2c reporter mRNAs. k Determination of editing windows for CU5.17 on C 459 of EGFP mRNA. Editing efficacy of CU5.17 on C 1541 of DDX3X reporter mRNA ( l ) and C 143 of SOD1 reporter mRNA ( m ) expressed in HEK293T cells. n Summary of the cytosine editing window mediated by CU5.17 around the target sites of EGFP, SOD1 and DDX3X reporter mRNAs. In a –n values represent mean ± SEM ( n = 3).

Article Snippet: Serum levels of PCSK9 were measured using the Mouse PCSK9 ELISA Kit (MPC-900, R&D Systems).

Techniques: Binding Assay, Sequencing

Journal: Nature Communications

Article Title: Effective in vivo RNA base editing via engineered cytidine deaminase APOBECs fused with PUF proteins

doi: 10.1038/s41467-025-64748-6

Figure Lengend Snippet: a Efficient editing of C 832 in PCSK9 mRNA by CU-REWIREs and Cas13-based editor. The ePUF10 binding site in PCSK9 is underlined in blue, with on-target site marked in red (top). Editing rates of C 832 are measured by Sanger sequencing with triplicates, values represent mean ± SEM. b Schematic illustration of CU5.21 vectors delivered into mice through intravenous injection of AAV8 virus (top) and the key experimental steps (bottom). c In vivo knockdown frequencies induced by AAV8-CU5.21. Unpaired two-sided Student’s t -test was used; statistical values represent the mean ± SEM ( n = 5). * P < 0.05, P = 0.0094. d –g The levels of serum PCSK9 protein (P = 0.0005), cholesterol (P LDL-C = 0.0026, P HDL-C = 0.0148, P Total-C = 0.0042), glyceride, and total protein levels in the mice injected with CU5.21. Unpaired two-sided Student’s t -test was used; statistical values represent the mean ± SEM ( n = 5). * P < 0.05. h Statistical analysis of weight changes in mice following administration in the CU5.21 and EGFP control. Values represent mean ± SEM ( n = 5). i Scatter plots showing transcriptome-wide C-to-U RNA base editing in liver samples treated with CU5.21 and EGFP control in mice. Values represent mean ± SD ( n = 3).

Article Snippet: Serum levels of PCSK9 were measured using the Mouse PCSK9 ELISA Kit (MPC-900, R&D Systems).

Techniques: Binding Assay, Sequencing, Injection, Virus, In Vivo, Knockdown, Control