SINV -1PRF Deep Mutational Scan

Here we are using an Aria II cell sorter to fractionate our cell library based on a ratio of mKate to eGFP. In this experiment we have a library of genetically modified HEK293T cells that each stably express a single mutation in the SINV structural polyprotein. mKate is downstream of the SINV structural polyprotein and in the -1 frame generating signal only in the event of a -1 programmed ribosomal frameshift. eGFP expresses off of an IRES segment to control for expression levels. After sorting each fraction of cells is harvested and gDNA is extracted for next generation sequencing. This experiment allows us to measure the -1PRF efficiency of thousands of SINV variants simultaneously.
Conclusion:
The data provides a heat map that indicates the frameshifting efficiency of each possible codon level mutation throughout the SINV structural polyprotein.