Randomized In-Frame Gene Fusions for Phenotype Diversity

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Solution Overview

Problem

Current methods lack a systematic approach to harness the function-generating capability of fusion genes or polypeptides for large-scale modification of biological organisms, limiting the creation of novel phenotypes and enhanced productivity in organisms used for production of materials and compounds.

Innovation Solution

A method involving the creation and use of systematic, randomized, large-scale in-frame gene fusions or polynucleotide fusions to disrupt or alter existing genetic or biochemical mechanisms, generating new phenotypes by introducing randomized in-frame fusion polynucleotides into an organism's genome, thereby increasing diversity and productivity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If traditional genetic engineering methods (gain-of-function approaches) are used to modify organisms, then specific functions can be introduced or enhanced, but the ability to systematically generate novel phenotypes and increase diversity is limited

Engineering Contradiction:
Improvephenotype diversityVSAvoidsystematic approach complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent combines multiple polynucleotides encoding different proteins into fusion polynucleotides, creating organisms with novel phenotypes by merging genetic elements from different sources into unified functional units that produce fusion proteins with new or enhanced characteristics

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The invention systematically varies parameters such as polynucleotide sequence combinations, protein fusion arrangements, and expression levels to generate diverse phenotypes, allowing high-throughput screening for optimal organismal traits including growth rate and productivity

Inventive Principle:
Principle #35Parameter changes

2Productivity

If mutagenesis is used to create genetic changes, then some gain-of-function changes may occur, but loss-of-function changes are more common and novel phenotypes are difficult to obtain systematically

Engineering Contradiction:
Improveyield enhancementVSAvoidgain-of-function frequency
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent performs preliminary design of fusion polynucleotides by selecting and combining specific polynucleotides that are likely to produce desirable gain-of-function effects, rather than relying on random mutagenesis, thereby increasing the probability of obtaining productive phenotypes before transformation

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The fusion polynucleotides are designed to combine multiple functional elements into single genetic constructs that can perform multiple functions simultaneously, increasing the likelihood of achieving enhanced productivity through versatile phenotypic changes

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Productivity

If polynucleotide fusions are created to alter phenotype, then new characteristics can be generated, but the scale and systematization of such modifications are currently insufficient

Engineering Contradiction:
Improveorganism productivityVSAvoidfusion creation scalability
Core Design Contradiction:
ProductivityVSEase of manufacture

Solution Approach 1:

The patent segments the genome into multiple polynucleotide units that can be independently characterized and then systematically recombined into fusion polynucleotides, enabling scalable and modular creation of genetic modifications that can be easily manufactured and propagated

Inventive Principle:
Principle #1Segmentation

Data Source

PatentEP2935582B1Compositions and methods for creating altered and improved cells and organisms
Publication Date: 2019.12.04 ZIELER HELGE
  • EP2935582B1 patent drawingFigure 1
  • EP2935582B1 patent drawingFigure 2A~2F
  • EP2935582B1 patent drawingFigure 3A~3D

AI summary

The present invention provides compositions comprising randomized in-frame fusion polynucleotides and methods for introducing them into a host organism to identify desirable phenotypic changes that disrupt or alter existing genetic or biochemical mechanisms or pathways, thus creating novel characteristics of the transformed organism. Methods for using the compositions for increasing diversity within populations of organisms are also presented.