CRISPR-Edited B Cells Co-Expressing Antibodies and Cargo Proteins

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

There is an unmet need for improved compositions and methods to controllably express therapeutic proteins in individuals, specifically for reversible expression of therapeutic proteins that can be activated or eliminated in response to specific antigens or anti-idiotypic antibodies.

Innovation Solution

Modified B cells are engineered using CRISPR-Cas systems to produce heterologous antibodies and co-express cargo proteins, with the ability to be activated by antigen binding and reduced or eliminated by anti-idiotypic antibodies, through homologous recombination and DNA repair templates introduced into the B cells' chromosomes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If modified B cells are engineered to co-express cargo proteins and heterologous antibodies, then therapeutic protein expression is achieved, but device complexity increases

Engineering Contradiction:

Improvetherapeutic protein expressionVSAvoiddevice complexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent combines the cargo protein expression cassette and heterologous antibody expression cassette into a single integrated construct that is introduced into B cells. This merging approach allows simultaneous co-expression of both therapeutic proteins from a unified genetic architecture, reducing the number of separate genetic modifications needed while achieving dual protein production.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The modified B cells are designed to perform multiple functions: they express the cargo protein therapeutically, produce heterologous antibodies for antigen recognition, and enable controllable activation or elimination mechanisms. This multi-functionality is achieved through a single genetic construct that coordinates expression of both proteins and incorporates regulatory elements for controlled cell fate.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Manufacturing precision

If CRISPR-Cas systems are used for chromosome cleavage and homologous recombination, then manufacturing precision is improved, but device complexity increases

Engineering Contradiction:

Improvechromosome integration precisionVSAvoiddevice complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent uses CRISPR-Cas systems as intermediary tools to facilitate precise chromosomal integration of the dual-expression construct. The guide RNA and Cas nuclease work together as a molecular intermediary system to locate specific genomic target sites and enable accurate insertion of the therapeutic construct through homology-directed repair, ensuring precise manufacturing without requiring complex manual genome editing procedures.

Inventive Principle:

Principle #24Intermediary (Mediator)

3Reliability

If anti-idiotypic antibodies are used to eliminate modified B cells, then reliability of controlled elimination is improved, but object-affected harmful factors increase

Engineering Contradiction:

Improvecontrolled cell eliminationVSAvoidimmune response effects

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent converts the potential harmful immune response against modified B cells into a beneficial control mechanism. By engineering the B cells to express heterologous antibodies with known idiotypes, the system enables specific anti-idiotypic antibodies to selectively target and eliminate only the modified B cells that express the therapeutic cargo, while leaving normal B cells unaffected. This transforms what could be an unwanted immune reaction into a precise suicide switch for controlled cell elimination.

Inventive Principle:

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

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 allows for conditional expression of therapeutic proteins, providing prophylactic and therapeutic benefits while maintaining allelic exclusion and safety, with efficient production and regulation of the expressed proteins.

Implementation Method 1

Methodology Applied

Scientific EffectCRISPR-Cas cleavage:

Implementation Method 2

The modified B cells are produced using homologous recombination that is facilitated in part using a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas (CRISPR-associated proteins) system for cleavage of the chromosome that allows homologous recombination between homologous segments of the chromosome and the repair templates

Methodology Applied

Scientific EffectHomologous recombination:

Data Source

PatentUS20230181636A1Antibody gene editing in b lymphocytes and co-expression of cargo protein

Publication Date: 2023.06.15 THE ROCKEFELLER UNIV

AI summary

This disclosure provides modified B cells which produce heterologous antibodies and co-express cargo proteins. The modified B cells may be stimulated by binding of a cognate antigen to the heterologous antibodies. The B cells may be reduced or eliminated by contacting the heterologous antibody with an anti-idiotypic antibody. Methods of making, and using the modified B cells for prophylaxis and therapy for a variety of conditions are provided. The B cells are modified at an IgH locus, an IgK locus, and combinations thereof. Modified B cells maintain allelic exclusion.