Engineered Effector Proteins for Precise Nucleic Acid Recognition

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current CRISPR/Cas systems for nucleic acid modification and detection face challenges in specificity and efficiency, particularly in targeting and modifying human genes associated with diseases, due to limitations in sequence recognition and cleavage capabilities.

Innovation Solution

Development of polypeptides and guide nucleic acids with engineered sequences that are at least 75% identical to specific sequences, combined with partner polypeptides and donor nucleic acids, to enhance target nucleic acid recognition and modification, including cis and trans cleavage activities, for precise editing and detection.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional CRISPR/Cas systems are used for nucleic acid modification, then the basic cleavage function is achieved, but the specificity and efficiency in targeting human genes are insufficient

Engineering Contradiction:
Improvetarget recognition specificityVSAvoidmodification efficiency
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The patent modifies the guide nucleic acid sequence parameters to achieve at least 75% identity with target sequences, and engineers effector proteins with customized amino acid sequences (at least 75% identical to Table 1 sequences) to enhance both specificity and cleavage efficiency for human gene targeting

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates composite systems combining engineered effector proteins with guide nucleic acids and partner polypeptides to achieve synergistic effects that improve both target recognition specificity and modification efficiency beyond what single components can achieve alone

Inventive Principle:
Principle #40Composite materials

2Productivity

If effector proteins with enhanced cleavage activity are used, then modification efficiency improves, but potential harmful effects on non-target sequences increase

Engineering Contradiction:
Improvecleavage efficiencyVSAvoidoff-target effects
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent designs guide nucleic acids with specific sequence compositions (at least 75% identity to targets) and engineered effector proteins with localized catalytic domains that work cooperatively to achieve high cleavage efficiency only at intended targets while minimizing off-target effects through precise local recognition

Inventive Principle:
Principle #3Local quality

3Measurement precision

If the system is designed for high specificity in target recognition, then off-target effects are reduced, but the complexity of the system increases

Engineering Contradiction:
Improvetarget recognition specificityVSAvoidsystem complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent divides the system into distinct functional modules: engineered effector proteins with specific catalytic domains, guide nucleic acids with target-complementary sequences, and partner polypeptides with coordinating functions. This segmentation allows each component to be optimized independently for specificity while maintaining overall system manageability

Inventive Principle:
Principle #1Segmentation

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 solution enables precise recognition and modification of target nucleic acids, including human genes, with improved specificity and efficiency, allowing for effective treatment of diseases associated with genetic mutations.

Implementation Method 1

CRISPR/Cas systems provide immunity in bacteria and archaea against viruses and plasmids by targeting the nucleic acids of the viruses and plasmids in a sequence-specific manner

Methodology Applied
Scientific EffectSequence-specific recognition:

Implementation Method 2

Nucleic acid modifying activities may include cis cleavage activity, trans cleavage activity, nicking activity, nuclease activity

Methodology Applied
Scientific EffectNuclease activity:

Data Source

PatentUS20240226327A9Effector proteins, effector partners, compositions, systems and methods of use thereof
Publication Date: 2024.07.11 MAMMOTH BIOSCIENCES INC

AI summary

Provided herein are compositions, systems, and methods comprising effector proteins, effector partners, and uses thereof. These effector proteins may be characterized as CRISPR-associated (Cas) proteins. Various compositions, systems, and methods of the present disclosure may leverage the activities of these effector proteins for the modification, detection, and/or engineering of nucleic acids.