Enhancing Prime Editing Efficiency Through Modified pegRNAs

Prime editing holds tremendous promise for precise genome engineering, but achieving efficient editing without increasing pegRNA complexity remains a significant challenge. Join this webinar to discover a novel pegRNA modification strategy that enhances stability and editing efficiency across multiple cell types without extending sequence length, providing actionable insights that can help accelerate prime editing research and streamline future development efforts.

Developing novel editing systems that do not require DSBs is an ever-expanding area of genetic research. Prime editing was developed as a “search-and-replace” genome editing technology. It comprises three key components:

1. Prime Editing Guide RNA (pegRNA)
2. Cas9 H840A Nickase
3. M-MLV Reverse Transcriptase

Despite its potential, the widespread application of prime editing has been limited by suboptimal editing efficiency. While studies have demonstrated that stabilizing the 3’ moiety through structural modifications (e.g., TeveproQ, G-quadruplex or hairpin motifs) can enhance efficiency, the elongation on sequence length might become problematic during CMC stage. As the general length for pegRNA would be around 130~150nt, it would be challenging to develop corresponding production processes and analytical methods.

To address this issue, GenScript screened novel modification combinations without elongating the sequence length to improve the editing efficiency. The optimized modification combination was found to be effective across multiple cell types, including HEK293T, HepG2, Jurkat and primary T cells. Further investigation revealed that the modification-optimized pegRNAs were found to be much more stable in the cytoplasm compared to canonical modified pegRNAs (three 2’-OMe and phosphorothioate on both ends).

Register now to learn how optimized pegRNA modifications can drive stronger editing outcomes in your research.

Speaker

Jacob Guo, PhD, Director of Nucleic Acid R&D Platform, GenScript

Dr. Jacob (Chun) Guo is the Director of the Nucleic Acid R&D Platform within the Life Science Group at GenScript. He earned his PhD in Chemistry from the University of Georgia, with expertise in nucleic acid chemistry and oligonucleotide modification.

Dr. Guo has authored five first-author publications in high-impact, peer-reviewed journals and has filed multiple patents related to oligonucleotide bioconjugation and their application in diagnostic technologies. He brings over 10 years of experience in solid-phase oligonucleotide synthesis. His recent work focuses on the manufacturing and quality control of synthetic long oligonucleotides for gene editing purposes and developing bio-conjugation methods for various oligonucleotide antibody/protein/peptide conjugates.

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