PROGRAMMABLE RNA WRITING USING CRISPR EFFECTORS AND TRANS-SPLICING TEMPLATESThis disclosure provides systems, methods, and compositions for site specific genetic engineering comprising the use of CRISPR effectors and trans-splicing. The disclosure also relates to methods of using the systems and compositions for treating diseases as ...
device in which Clover fluorescence surpasses mCherry fluorescence. Analyzing devices harboring modified PPT elements using long-read sequencing shows significant changes to splicing, with an aberrant double inclusion RNA product being dominant. Of these devices, the secondary RNA product from the device ...
Finally, mRNA was purified using the RNA Clean and Concentrator-5 kit (Zymo). Transient transfection On the day before transfection, HEK cells were seeded at a density of 0.3-0.6 million cells per well on a 12-well plate, 0.1-0.3 million cells per well on a 24-well plate, or 15,000...
We developed luciferase-based splicing reporters to evaluate 718 RNA-binding proteins (RBPs) for the ability to activate exon inclusion. Our screen detected RBPs with no previously characterized roles in splicing and utilized RBP domains displaying potent exon activation to develop programmable splicing...