Results are obtained that reveal generation of an RNA/DNA hybrid in yeast chromosomal DNA based on the transfer of genetic information from the RNA tract of the oligo to a mutated marker gene present in the yeast genome. As shown in this video, transformation using RNA-containing oligos requires essentially only a few additional steps compared to transformation using DNA molecules, to prevent degradation by RNases. Thus, transformation using RNA-containing oligos is not limited to the yeast system, but can be applied to any organism or cell type where transformation by DNA oligos is proficient.

While several in vitro studies have been conducted to characterize the function of factors that can recognize RNA in a hybrid with DNA, the identity and the in vivo functions of factors that could affect RNA/DNA hybrid stability remain mostly unknown. The possibility to utilize RNA-containing oligos of a significant length (50 to 80-mers) and optimal quality (such as Thermo Scientific Dharmacon RNA-containing oligos) has opened the way to examining a wide range of molecular processes directly in vivo in the cells of interest. As shown in this video, targeting cells by RNA-containing oligos provides the opportunity to embed ribonucleotides into genomic DNA in cells. The stability, function and consequences of these in vivo generated RNA/DNA hybrids can be analyzed and characterized, potentially uncovering unknown mechanisms of RNA-driven DNA editing and modification.

The work was supported by the Georgia Cancer Coalition grant-R9028.
The video production for this article was sponsored by Thermo Fisher Scientific.