Engineered Transposase Variants for Targeted Genome Integration

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Solution Overview

Problem

Existing transposon systems, such as the Sleeping Beauty (SB) transposon, exhibit random integration into the genome, which poses a genotoxic risk, particularly in human applications, due to potential oncogenic transformation.

Innovation Solution

Development of specific transposase variants with enhanced specificity of integration into the genome, particularly into palindromic AT repeat target sequences, which reduces integration into exons and transcriptional regulatory regions, thereby enhancing safety and utility in gene therapy applications.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If random integration into the genome is used, then transpositional activity is high, but genotoxic risk increases due to potential oncogenic transformation

Engineering Contradiction:

Improvetranspositional activityVSAvoidgenotoxic risk

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The transposase variants exhibit altered local properties by showing preferential integration into specific genomic regions (heterochromatin, repetitive sequences, intergenic regions) while maintaining overall transpositional activity. This local targeting quality reduces genotoxic risk without sacrificing productivity

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

Amino acid substitutions in the transposase variants (e.g., H187V, P247R, K248R) change the molecular parameters of the enzyme, leading to altered integration specificity. These parameter changes redirect integration away from dangerous regions while preserving transpositional efficiency

Inventive Principle:

Principle #35Parameter changes

2Productivity

If integration into exons and transcriptional regulatory regions is increased, then gene transfer efficiency is high, but safety is reduced due to potential disruption of essential gene functions

Engineering Contradiction:

Improvegene transfer efficiencyVSAvoidsafety

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The invention converts the potential harm of random integration into a benefit by directing integration toward safe genomic regions. The transposase variants use their altered specificity to target heterochromatin and repetitive sequences, transforming what could be dangerous random insertion into a safe and predictable integration pattern that preserves gene function

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

Data Source

PatentUS20250129389A1Polypeptides with transpositional activity

Publication Date: 2025.04.24 MAX DELBRUECK CENT FUER MOLEKULARE MEDIZIN

AI summary

The present invention relates to polypeptides with transpositional activity, particularly engineered polypeptides with transpositional activity, nucleic acids encoding such polypeptides, vectors compris-ing said nucleic acids, a cell comprising said nucleic acid, methods of integrating an exogenous nucle-ic acid into the genome using said polypeptides, said polypeptides for use in medicine and/or several gene therapies; and a pharmaceutical composition comprising said polypeptides, nucleic acids, vectors or cells. Particularly, the present invention relates to said polypeptide, which is engineered to have a gain of specificity of integration into the genome.