Genomic Insertion of Large Cargo with Engineered Retro Elements

Technology

The system includes an R2 element enzyme or a similar non-LTR site specific retrotransposon element, and a payload RNA with an insertion region, optional homology region, and a protein binding element for insertion. The enzyme may have a natural or engineered targeting domain that directs it to the specific site in the genome, which can be a genomic locus other than the 28S rRNA site. The payload RNA, which includes a template for the genetic insertion, is introduced into the genome of a specific cell type, and in some embodiments, can function in post-mitotic cells. The enzyme’s reverse transcriptase domain converts the RNA template into DNA, and the nickase domain facilitates the insertion of this DNA into the genome.

Problem Addressed

This technology significantly reduces off-target effects and enhances the success rate of genetic edits, allowing applications to more complex genomic corrections. Unlike traditional methods which may struggle with accurately inserting large genetic elements, this system can carry RNA molecules of varying sizes and is capable of directed insertions. Additionally, the R2 element enzyme can be modified with natural or engineered targeting domains, zinc finger motifs, or fusion with other proteins like Cas9 or TALEN, enabling more accurate control of the insertion process. Overall, this technology overcomes the limitations of size, cargo capacity, and targeting ability found in existing genome editing systems.

Advantages

• Enables targeted insertion at specific genome loci
• Capable of inserting large genetic elements effectively
• Ensures precise and controlled insertion of genetic material
• Versatile applications, including programmable genetic corrections for diseases like cystic fibrosis