Type I-C CRISPR RNP for Compact Mammalian Genome Editing

Solution Overview

Problem

Existing type I-E CRISPR systems for genome editing in mammalian cells are cumbersome due to their large size and complex composition, making delivery challenging and inefficient, and they induce heterogeneous, large DNA segment deletions that are difficult to manage.

Innovation Solution

Utilizing a compact type I-C CRISPR system from Neisseria lactamica, comprising only four Cas genes and a CRISPR array, which forms a streamlined ribonucleoprotein (RNP) for efficient genome editing in human cells, inducing unidirectional, heterogeneous large deletions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If type I-E CRISPR-Cas system is used for genome editing, then genome editing function is achieved, but gene size is large and delivery into mammalian cells is difficult

Engineering Contradiction:

Improvegenome editing functionVSAvoidgene size

Core Design Contradiction:

ReliabilityVSVolume of moving object

Solution Approach 1:

The type I-E CRISPR-Cas system is divided into separate functional modules: Cascade complex (Cas5, Cas8, Cas7, crRNA) and Cas3 endonuclease. This segmentation allows each component to be optimized and delivered separately, reducing the burden of delivering the entire system as one large construct.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention extracts and utilizes only the essential components for genome editing function (Cascade complex and Cas3), removing non-essential elements. The streamlined type I-C system from Neisseria lactamica requires only four Cas genes and one CRISPR array, eliminating redundant genetic material while maintaining editing capability.

Inventive Principle:

Principle #2Taking out (Extraction)

2Reliability

If type I-E CRISPR-Cas system is used for genome editing, then genome editing function is achieved, but ribonucleotide preparation is inefficient

Engineering Contradiction:

Improvegenome editing functionVSAvoidribonucleotide preparation efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The invention extracts and utilizes only the essential components for genome editing function (Cascade complex and Cas3), removing non-essential elements. The streamlined type I-C system from Neisseria lactamica requires only four Cas genes and one CRISPR array, eliminating redundant genetic material while maintaining editing capability.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the system parameters by switching from type I-E to type I-C CRISPR-Cas system, which has inherently fewer components and smaller gene size. This parameter change fundamentally improves preparation efficiency while maintaining editing function.

Inventive Principle:

Principle #35Parameter changes

3Ease of manufacture

If type I-C CRISPR-Cas system with fewer Cas genes is used, then delivery efficiency and preparation ease are improved, but genome editing activity must be maintained

Engineering Contradiction:

Improvepreparation easeVSAvoidgenome editing activity

Core Design Contradiction:

Ease of manufactureVSReliability

Solution Approach 1:

The invention changes the system parameters by switching from type I-E to type I-C CRISPR-Cas system, which has inherently fewer components and smaller gene size. This parameter change fundamentally improves preparation efficiency while maintaining editing function.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention uses a simplified copy of the CRISPR system (type I-C instead of type I-E) that retains the essential editing function while removing redundant components. The core mechanism is copied and adapted to a more efficient system.

Inventive Principle:

Principle #26Copying

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

The Nla type I-C CRISPR system achieves high genome editing activity (50-96%) in human cells, facilitating large-scale production and delivery via viral vectors or mRNA, with robust and programmable long-range deletions.

Implementation Method 1

a synthetic guide RNA sequence that is complementary to a target genomic DNA sequence in a host cell

Methodology Applied

Scientific EffectComplementary base pairing:

Implementation Method 2

the combination of two or more N. lactamica proteins induces cleavage of one or both strands in the target DNA sequence, thereby altering the target DNA sequence

Methodology Applied

Scientific EffectDNA cleavage:

Data Source

PatentUS12630846B2Type I-C CRISPR system from <i>Neisseria lactamica </i>and methods of use

Publication Date: 2026.05.19 CORNELL UNIVERSITY

AI summary

The disclosure provides a method of altering a DNA sequence, which method comprises the use of a type I-C CRISPR-Cas system based on Neisseria lactamica. The system involves a combination of two or more Neisseria lactamica proteins selected from Cas3, Cas5, Cas8c, and Cas7 and a synthetic guide RNA sequence.