Segmented T7 Promoter Primers for Nucleic Acid Amplification

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Solution Overview

Problem

Existing templated nucleic acid synthesis technologies are limited in terms of speed, efficiency, and quality of product generation.

Innovation Solution

A method involving the use of primers with T7 promoter sequence elements and CRISPR-Cas detection composition for amplifying and detecting nucleic acid sequences, including a guide polynucleotide, labeled nucleic acid reporter construct, and Cas protein, such as Cas13 or Cas12, to enhance nucleic acid production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional templated nucleic acid synthesis is used, then the process is simple, but the speed and efficiency of product generation are limited

Engineering Contradiction:

Improvespeed and efficiency of nucleic acid product generationVSAvoidcomplexity of amplification system

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The primer is segmented into distinct functional elements: a T7 promoter sequence element and a hybridization element. This segmentation allows each element to perform its specific function independently, with the promoter element driving high-speed transcription and the hybridization element ensuring specific binding to the target sequence, thereby resolving the contradiction between speed and complexity.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The primer serves multiple functions simultaneously: it acts as both a binding element (via the hybridization element) and a transcriptional driver (via the T7 promoter element). This multi-functionality consolidates what would otherwise require separate components into a single reagent, improving productivity without proportionally increasing system complexity.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Manufacturing precision

If conventional primers are used, then the system is simple, but the quality of nucleic acid amplification is limited

Engineering Contradiction:

Improvequality of nucleic acid amplificationVSAvoidcomplexity of primer design

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The primer is divided into functionally distinct segments: a T7 promoter sequence element that ensures high-quality transcription initiation and a hybridization element that provides specific sequence binding. This segmentation allows optimization of each function independently, improving amplification quality without requiring complex multi-component systems.

Inventive Principle:

Principle #1Segmentation

3Manufacturing precision

If T7 promoter sequence elements are added to primers, then nucleic acid amplification quality improves, but the primer design complexity increases

Engineering Contradiction:

Improvequality of nucleic acid amplificationVSAvoidprimer design complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The primer is segmented into a T7 promoter sequence element and a hybridization element, allowing each segment to be designed and optimized independently. The promoter element is standardized to ensure consistent transcriptional activity, while the hybridization element is tailored for specific target binding, thereby improving quality without requiring complex overall design.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The invention changes the sequence parameters of the primer by incorporating the T7 promoter motif (specific nucleotide sequence) while maintaining the hybridization element's complementarity to the target. This parameter change enables high-quality amplification through standardized promoter recognition by T7 polymerase, avoiding the need for complex custom designs.

Inventive Principle:

Principle #35Parameter changes

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

Improves the speed, efficiency, and quality of nucleic acid amplification and detection processes.

Implementation Method 1

transcribing a copied and/or amplified templated nucleic acid synthesis target using any primer inserted promoter

Methodology Applied

Scientific EffectTranscription:

Implementation Method 2

contacting the amplified nucleic acid comprising the target sequence of interest with a CRISPR-Cas detection composition; and detecting the amplified nucleic acid

Methodology Applied

Scientific EffectCRISPR-Cas detection: