GeneRulor Secures Six Abstracts at ASGCT 2026

The American Society of Gene and Cell Therapy (ASGCT) 2026 Annual Meeting will be held in Boston, USA, from May 11 to 15, 2026.

GeneRulor has six research abstracts accepted for poster presentation, demonstrating its systematic capabilities and technological

advances in genome editing tool development and genomic safety assessment.

The six selected studies represent not isolated technical advances, but a comprehensive technology framework spanning the full

workflow of genome editing therapeutics—from tool development and optimization to off-target detection and genome-wide

safety evaluation—forming a closed-loop platform.

Title：Ino-seq enables comprehensive profiling of adenine base editor off-targets

Adenine base editors (ABEs) have entered clinical development; however, existing off-target detection methods exhibit significant limitations. GUIDE-seq and CHANGE-seq-BE fail to capture sgRNA-independent

off-target events, while whole-genome sequencing (WGS) shows extremely low sensitivity (F1-score < 0.01).

GeneRulor developed Ino-seq, which leverages endonuclease V to directly cleave inosine-containing DNA—the immediate product of

ABE editing—enabling unbiased and high-resolution detection of off-target sites.

Across eight therapeutic targets, Ino-seq achieved an F1-score of 0.892-0.931, with a validation accuracy exceeding 95.3% (1,330/1,396 sites validated), significantly outperforming WGS (0.001–0.006) and GUIDE-seq (0.043–0.213).

Importantly, Ino-seq identified 892 off-target sites missed by all other methods, including 47 sites with editing frequencies >5%,

which may represent clinically relevant safety risks. These findings establish a new technical benchmark for genotoxicity assessment in IND-enabling studies.

Title：LongTn-seq: A Novel Long-Read Tn5-Based Sequencing Platform for Comprehensive Quantification of AAV-Mediated HDR Efficiency and Genomic Safety Assessment

AAV-mediated homology-directed repair (HDR) is a key strategy for precise gene insertion, yet conventional methods cannot simultaneously evaluate integration efficiency and genomic safety.

GeneRulor developed LongTn-seq, integrating Tn5-based library construction with long-read sequencing (PacBio), enabling simultaneous quantification of HDR efficiency, insertion fidelity, and chromosomal rearrangements within a single assay.

Validation experiments showed an AAV-HDR integration rate of 72.29%, alongside detailed characterization of editing outcomes, including small deletions (3.59%), large deletions (1.32%), insertions (1.11%), and inversions (0.02%). Interchromosomal translocation events were rare (0.05%, n=3), indicating high targeting specificity.

This represents the first platform capable of comprehensive HDR quantification and concurrent genomic safety assessment in a single experiment, directly addressing regulatory expectations for genome integrity evaluation.

Title：Plug-in Base Editor: A Modular Platform for Programmable Control of Editing Windows, Efficiency, and Fidelity in DNA Base Editors

Conventional base editors position deaminases at fixed N- or C-termini of Cas9, resulting in constrained editing windows (typically positions 4–8), limiting correction of ~50%–75% of clinically relevant pathogenic variants.

GeneRulor developed the Plug-in Base Editor system, which enables dynamic repositioning of deaminases via epitope–antibody interactions.

A total of 62 epitope insertion sites were engineered on nCas9, and 533 modular combinations were systematically evaluated. The system increased C-to-T editing efficiency to ~90% while reducing byproduct formation from 25%–37% to 2%–4%.

In vivo validation in zebrafish demonstrated a 3.8-fold increase in editing efficiency at the TWIST2 locus, with precise single-base editing rates increasing from 10% to 59%.

ClinVar analysis further showed expansion of correctable variants: A-to-G variants increased from 23.26% to 32.87%, and C-to-T variants from 43.53% to 77.73%.

Title：Systematic high-throughput evaluation reveals FrCas9’s superior specificity and efficiency for therapeutic genome editing

While SpCas9 is widely used, off-target activity remains a major safety concern. GeneRulor established a three-stage high-throughput screening pipeline (AID-seq → amplicon sequencing → GUIDE-seq validation) to evaluate FrCas9 derived from Faecalibaculum rodentium.

Across 21 therapeutically relevant targets (including AAVS1, B2M, CD7, PDCD1, TRAC), FrCas9 achieved editing efficiencies exceeding 70% at multiple loci (e.g., hROSA26: 91.3%, B2M: 80.9%).

Compared to SpCas9 and OpenCRISPR-1, FrCas9 exhibited significantly reduced off-target activity. Fusion with TREX2 further improved genome stability by reducing large deletions and chromosomal translocations.

Title：Structural and functional bases of F. rodentium Cas9 provide insights into CRISPR-Cas protein engineering

Cryo-EM analysis resolved the FrCas9–sgRNA–DNA complex at 2.89 Å resolution, revealing two functional states: the R-loop expansion state and the pre-catalytic state.

Structural analysis identified unique over-twisting of the sgRNA–DNA heteroduplex and a phospho-lock loop (PLL)-mediated mismatch sensing mechanism, providing a molecular basis for enhanced specificity.

Structure-guided engineering yielded an improved variant, eFrCas9, which maintains on-target activity while reducing off-target effects. Therapeutic potential was validated in a Duchenne muscular dystrophy (DMD) cell model.

Title：Methodological Validation of Liquid-Phase Hybrid Capture for AAV Vector Integration Site Detection

Although AAV integration occurs at low frequency, both FDA and EMA require comprehensive integration site analysis.

Conventional methods (nrLAM-seq, LM-seq) focus on ITR regions and may miss non-ITR integration events.

GeneRulor validated a liquid-phase hybrid capture approach using probes covering the full AAV genome. Two integration sites were detected and confirmed by Sanger sequencing, whereas conventional methods failed to identify these events.

Dilution experiments demonstrated a detection limit of 0.001%, meeting regulatory requirements for preclinical and clinical development.

These studies collectively demonstrate GeneRulor’s integrated approach:

Editing tools: Plug-in BE and FrCas9 expand editing capability and specificity

Safety assessment: Ino-seq and LongTn-seq enable comprehensive genomic evaluation

Analytical methods: Hybrid capture improves integration site detection sensitivity

This platform provides a quantitative, validated, and regulatory-aligned framework to support the clinical translation of genome editing therapeutics.