| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | XTEN and COM inserts from pN22p-UGI and pUC57-COM (synthesized by GenScript), respectively. Then, NLS-UGI, XTEN and COM inserts and the linear pMMLVgag(C507V)-3×NES vector were recombined to generate the pMMLVgag(C507V)-UGI-COM. Cells and VLPs were lysed in protein loading buffer (TransGen Biotech, DL101) and incubated at 95 °C for 20 min, before being separated by SDS–PAGE (Genscript, M00661) in loading buffer. Modified sgRNA for mouse studies was synthesized by GenScript. | Get A Quote |
| Synthetic Guide RNA | Get A Quote |
Virus-like particles (VLPs) are promising for delivering genome editors, yet the in vivo in vivo efficacy of VLP-mediated cytosine base editing remains limited. Here we identified insufficient inhibition of uracil DNA glycosylases as the underlying mechanism of low cytosine base editor (CBE) editing efficiencies in vivo. We engineered a previously reported CBE, transformer base editor (tBE), and developed a VLP delivery system to enhance the recruitment of uracil DNA glycosylase inhibitor proteins. tBE-VLPs achieved robust C-to-T editing in mouse liver and retina. A single injection achieved, on average, 46.0% editing at mPcsk9 and 64.2% at mHpd in the liver, as well as 24.2% at mVegfa in the retinal pigment ep... More
