PhD Scientific Days 2026

Poster Session 2.C - Molecular Medicine

ProPE Expands the Prime Editing Window and Enhances Gene Editing Efficiency Where Prime Editing is Inefficient

Name of the presenter

Simon, Dorottya Anna

Institute/workplace of the presenter

HUN-REN TTK

Authors

Dorottya Anna Simon^1,2, Sarah Laura Krausz^1,2, Zsuzsa Bartos¹, Zsuzsanna Biczók^1,2, Éva Varga^1,3, Krisztina Huszár¹, Péter István Kulcsár¹, András Tálas¹, Zoltán Ligeti¹, Ervin Welker¹
1: Institute of Molecular Life Sciences, HUN-REN Research Centre for Natural Sciences, Budapest, Hungary
2: School of PhD Studies, Semmelweis University, Budapest, Hungary
3: School of PhD Studies, University of Szeged, Szeged, Hungary

Text of the abstract

Introduction: Prime editing (PE) is a promising gene editing method that exploits a reverse transcriptase fused to a Cas9, whose single guide RNA (sgRNA) is extended with a reverse transcriptase template containing the desired DNA modifications. Its efficiency and specificity are inconsistent, requiring extensive optimization.
Aims: To develop a prime editing tool, which overcomes multiple key limitations of PE activity, and to assess its therapeutic potential.
Methods: molecular cloning; cell culturing and transfection (HEK293, K562, U2OS, HuES9); flow cytometry; next-generation sequencing
Results: We propose prime editing with prolonged editing window (proPE), which uses a second non-cleaving sgRNA to target the reverse transcriptase template near the edit site. ProPE requires less optimization than PE and extends PE’s potential for allele-specific modifications. By overcoming five limitations of PE, proPE significantly increases overall editing efficiency 6.2-fold up to 29.3% for low-performing edits (<5% with PE) and broadens its applicability to modifications beyond the typical PE range.
Conclusions: By increasing the editing efficiency and range, proPE may make it practical to generate disease models for the majority of pathogenic single nucleotide polymorphisms and may also considerably contribute to expanding the potential of PE to enable its use in therapeutic interventions.
Funding: Support was provided by the Ministry of Culture and Innovation of Hungary through the National Research, Development and Innovation Fund (project no. KDP-12-10/PALY-2022), financed under the KDP-2021 funding scheme.
Email: simon.dorottya@ttk.hu, University: Semmelweis University, Supervisor: Ervin Welker