PFTα and bFGF Mediators for Gene Editing Efficiency

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Solution Overview

Problem

Current genome editing methods, such as CRISPR/Cas9, often result in DNA damage and apoptosis in cells, limiting gene editing efficiency and survival rates, particularly in cells intended for xenotransplantation like porcine cells.

Innovation Solution

The use of exogenous PFTα or bFGF during genetic modification in cells, such as porcine cells, to inhibit apoptosis and enhance gene editing efficiency by reducing DNA damage and promoting cell survival.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If CRISPR/Cas9 nuclease is used to edit genomes in cells, then gene editing efficiency is improved, but DNA damage and apoptosis increase

Engineering Contradiction:

Improvegene editing efficiencyVSAvoidDNA damage and apoptosis

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent introduces an intermediary substance (exogenous PFTα or bFGF) that mediates between the harmful DNA damage caused by CRISPR/Cas9 and the cell's survival mechanisms. This intermediary inhibits apoptosis and promotes cell survival, allowing the beneficial gene editing effect to prevail while mitigating the harmful effects.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the biological parameters of the cell by introducing exogenous factors (PFTα or bFGF) that alter the cell's response to DNA damage. These parameter changes enable the cell to withstand the stress of nuclease-mediated editing while maintaining editing efficiency and improving survival rates.

Inventive Principle:

Principle #35Parameter changes

2Manufacturing precision

If site-specific nucleases are delivered to generate DNA double strand breaks, then gene targeting is achieved, but cell survival rates decrease

Engineering Contradiction:

Improvegene targeting accuracyVSAvoidcell survival rate

Core Design Contradiction:

Manufacturing precisionVSReliability

Solution Approach 1:

The exogenous PFTα or bFGF acts as a protective intermediary that shields the cell from the lethal effects of DNA double strand breaks while allowing the targeted gene editing to proceed. This mediator enables precise gene targeting without the corresponding decrease in cell survival.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent applies beforehand cushioning by introducing exogenous PFTα or bFGF before or during the nuclease treatment to preemptively protect the cell from apoptosis. This protective measure is established in advance to cushion against the harmful effects of DNA damage, ensuring cell survival while maintaining gene targeting accuracy.

Inventive Principle:

Principle #11Beforehand cushioning (Prior cushioning)

3Productivity

If multiple target nucleic acid sequences are broken in a cell, then gene editing efficiency increases, but apoptosis risk increases

Engineering Contradiction:

Improvegene editing efficiencyVSAvoidapoptosis risk

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The exogenous PFTα or bFGF serves as a protective intermediary that allows multiple nucleic acid sequences to be targeted and broken simultaneously while mitigating the increased apoptosis risk. This mediator enables high productivity gene editing without the proportional increase in cell death.

Inventive Principle:

Principle #24Intermediary (Mediator)

Data Source

PatentUS12325865B2Methods for increasing efficiency of gene editing in cells

Publication Date: 2025.06.10 PRESIDENT & FELLOWS OF HARVARD COLLEGE

AI summary

The present disclosure provides methods of improving nuclease-mediated gene targeting frequencies in cells using PFTα or bFGF.