Targeted Enveloped CRISPR Delivery to Limit Off-Target Editing

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current delivery strategies for CRISPR-Cas effector polypeptides result in prolonged expression in target cells, increasing the likelihood of off-target gene editing events.

Innovation Solution

Development of enveloped delivery vehicles (EDVs) comprising a viral envelope protein and a targeting polypeptide for specific delivery of CRISPR-Cas effector polypeptides to target cells, utilizing fusion proteins and guide RNAs for precise genome editing.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If a recombinant virus encoding a CRISPR-Cas effector polypeptide is used for delivery, then the CRISPR-Cas effector polypeptide is delivered into target cells, but prolonged expression occurs which increases the likelihood of off-target gene editing events

Engineering Contradiction:

Improvespecificity of genome editingVSAvoidexpression duration of CRISPR-Cas effector polypeptide

Core Design Contradiction:

ReliabilityVSDuration of action of moving object

Solution Approach 1:

The delivery system is segmented into multiple functional components: an enveloped virus-like particle structure for delivery, a CRISPR-Cas effector polypeptide for genome editing, and a guide RNA for target recognition. This segmentation allows independent optimization of each component's function and duration of action.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The CRISPR-Cas effector polypeptide is pre-assembled with the guide RNA to form a ribonucleoprotein complex before delivery. This preliminary assembly ensures that the effector polypeptide is immediately active upon delivery and reduces the time required for intracellular expression and assembly, thereby limiting prolonged expression.

Inventive Principle:

Principle #10Preliminary action

2Adaptability or versatility

If enveloped delivery vehicles with targeting polypeptides are used, then targeted delivery to specific cell types is achieved, but the device complexity increases

Engineering Contradiction:

Improvetargeting specificity to cell typesVSAvoidstructure of delivery vehicle

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The enveloped virus-like particle structure serves multiple functions: it provides protective encapsulation for the CRISPR-Cas effector polypeptide and guide RNA, facilitates cellular entry through viral fusion mechanisms, and enables targeted delivery when equipped with cell-type-specific envelope proteins. This multi-functionality reduces the need for separate delivery systems for different cell types.

Inventive Principle:

Principle #6Universality (Multi-functionality)

Solution Approach 2:

Cell-type-specific envelope proteins act as intermediaries that mediate the interaction between the delivery vehicle and target cells. These envelope proteins recognize and bind to specific receptors on the surface of target cells, enabling selective delivery without requiring complex internal targeting mechanisms within the particle structure.

Inventive Principle:

Principle #24Intermediary (Mediator)

Data Source

PatentUS20260048143A1Compositions and Methods for Targeted Delivery of CRISPR-CAS Effector Polypeptides

Publication Date: 2026.02.19 RGT UNIV OF CALIFORNIA

AI summary

The present disclosure provides enveloped delivery vehicles (EDVs) comprising a nucleic acid-binding effector polypeptide, or a nucleic acid encoding the nucleic acid-binding effector polypeptide, where the EDV comprises a fusion polypeptide comprising (i) a viral envelope protein and (ii) a targeting polypeptide that provides for binding to a target cell. The present disclosure provides methods of using an EDV of the present disclosure for delivery of, e.g., a nucleic acid-binding effector polypeptide, to a eukaryotic cell.