B2M Epigenetic Editors for Durable Gene Silencing Without DNA Breaks

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Traditional genetic engineering strategies for immune cells, such as those used in adoptive cell therapy, involve permanent genomic manipulations that can lead to risks like chromosomal translocations, nucleotide insertions, and off-target mutations, necessitating safer and more efficient methods for epigenetic modification.

Innovation Solution

The use of epigenetic editors, comprising fusion proteins or nucleic acid molecules with DNA methyltransferase (DNMT) domains, transcriptional repressor domains, and DNA-binding domains, specifically targeting the B2M gene in human cells to repress transcription without causing DNA breaks, utilizing CRISPR Cas, ZFP, or TALE domains for precise epigenetic regulation.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Duration of action of stationary object

If traditional genetic engineering strategies are used to permanently manipulate cells at the genomic level, then durable modification of gene expression is achieved, but risks including chromosomal translocations, undesired insertions and deletions of nucleotides, and off-target mutations occur

Engineering Contradiction:

Improvedurability of gene modificationVSAvoidchromosomal translocations and off-target mutations

Core Design Contradiction:

Duration of action of stationary objectVSObject-affected harmful factors

Solution Approach 1:

The patent changes the fundamental parameter of gene modification from permanent genomic alteration to reversible epigenetic modification. By using epigenetic editors to modify chromatin structure and DNA methylation states rather than altering the genomic sequence, the system achieves durable gene silencing while avoiding the harmful effects of permanent genetic manipulation such as chromosomal translocations and off-target mutations

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent introduces epigenetic editors as intermediary molecules that mediate between the desired gene silencing outcome and the genomic DNA. These editors work through intermediate mechanisms including DNA methylation and chromatin remodeling, allowing durable modification without directly altering the genomic sequence, thus avoiding harmful effects like chromosomal translocations

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If epigenetic editing is used to repress B2M transcription, then reversible and durable silencing is achieved without DNA breaks, but the system complexity increases due to multiple domain requirements

Engineering Contradiction:

Improvesafety profile without DNA breaksVSAvoidfusion protein domain structure

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent merges multiple functional domains into a single fusion protein structure. The epigenetic editor combines a DNA-binding domain (such as dCas9, ZFP, or TALE), a DNMT domain for DNA methylation, and a transcriptional repressor domain. This merging approach achieves reliable gene silencing without DNA breaks while managing system complexity through integrated protein design

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The fusion protein serves multiple functions simultaneously: the DNA-binding domain provides target specificity, the DNMT domain enables epigenetic modification through DNA methylation, and the transcriptional repressor domain ensures durable gene silencing. This multi-functionality in a single protein reduces the need for separate components and manages overall system complexity

Inventive Principle:

Principle #6Universality (Multi-functionality)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach provides reversible and durable silencing of the B2M gene, reducing alloreactivity in allogeneic cells with lower risks of chromosomal instability and off-target effects, offering a safer alternative to traditional genome editing.

Implementation Method 1

a DNA methyltransferase (DNMT) domain and/or a domain that recruits a DNMT

Methodology Applied

Scientific EffectDNA methylation:

Implementation Method 2

the DNA-binding domain comprises a dead CRISPR Cas (dCas) domain

Methodology Applied

Scientific EffectCRISPR Cas binding:

Implementation Method 3

a transcriptional repressor domain

Methodology Applied

Scientific EffectTranscriptional repression:

Data Source

PatentUS20250387518A1Compositions and methods for epigenetic regulation of b2m expression

Publication Date: 2025.12.25 NCHROMA BIO

AI summary

Disclosed herein are compositions and methods comprising epigenetic editors for epigenetic modification of B2M, as well as nucleic acids and vectors encoding the same. Also disclosed are cells epigenetically modified by the epigenetic editors.