Enhancing HDR Efficiency with Modified DNA Donors

Example 9

Modified DNA donors with a 5′ spermine modification were tested for their effect on HDR efficiency. Using the HE293 TLR assay, it was found that these spermine-modified DNA donors were more effective than unmodified DNA donors. The improvement in efficiency is comparable to that of TEG alone as well. When combining TEG and spermine modifications in the same DNA donor, there was also an increase in the HDR efficiency compared to the unmodified donors (FIG. 26).

These combined results indicate that 2′-OMe RNA, PEG, spermine, and combinations of these modifications make DNA donor molecules more potent for HDR.