Optically Encoded Particles for Nucleic Acid Screening
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for screening large libraries of edited cells are labor-intensive and time-consuming, requiring weeks to isolate and validate clonal cell lines, limiting the assessment of genetic interactions and cellular processes.
Innovation Solution
The use of optically encoded particles to deliver primers for nucleic acid amplification in discrete volumes, allowing for the identification and correlation of nucleic acid sequence variations with observable characteristics through the detection of unique particle combinations and subsequent sequencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional methods of picking individual cells and culturing in multi-well plates are used, then clonal cell lines can be isolated and validated, but the process requires two weeks of labor per isogenic line
Solution Approach 1:
The patent segments the screening process by distributing unique optically encoded particles into discrete volumes, each containing single cells or small cell groups. This segmentation allows parallel processing of thousands of discrete volumes simultaneously, reducing the time required from weeks to days while maintaining the precision of clonal cell line isolation and validation
Solution Approach 2:
The patent replaces the manual mechanical process of picking individual cells with an optical detection system. Optically encoded particles are detected using optical methods (fluorescence, absorbance, scattering), and the information is processed computationally to identify discrete volumes containing target cells, eliminating the labor-intensive manual picking process
2Adaptability or versatility
If large libraries of edited cells are screened using traditional methods, then genetic interactions can be assessed, but the cost and time requirements are prohibitively high
Solution Approach 1:
The patent creates a universal screening platform where optically encoded particles serve multiple functions: they act as barcodes for identification, deliver primers for nucleic acid amplification, and enable optical detection. This multi-functionality allows the same system to screen large libraries of edited cells across different genetic backgrounds and conditions, significantly improving productivity while reducing costs
Solution Approach 2:
The patent changes the parameters of the screening system by using optical detection parameters (fluorescence intensity, absorbance, scattering) instead of traditional phenotypic selection parameters. This allows for high-throughput screening of large cell libraries with improved productivity, as optical measurements can be automated and processed rapidly
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
This approach enables high-throughput, cost-effective, and rapid screening of nucleic acid sequence variations, reducing the time and labor required for isolating clonal cell lines and correlating sequence information with phenotypic assessments.
Implementation Method 1
A single solution comprising a set of optically encoded particles is then dispensed across all discrete volumes resulting in delivery of a combination of optically encoded particles to each discrete volume
Implementation Method 2
The combination of optically encoded particles is detected in each discrete volume thereby allowing each discrete volume to be identified by the observed combination of optically encoded particles
Implementation Method 3
Amplicons comprising the one or more target sequences are generated in each discrete volume using the one or more primers delivered to each discrete volume
Data Source
Figure 1
Figure 2A~2B
Figure 3
AI summary
Embodiments disclosed herein provide reagents and methods for high-throughput screening of nucleic acid sequence variations in nucleic acid containing specimens. Nucleic acid specimens to be screened are loaded into separate discrete volumes. Optically encoded particles are used to deliver primers to amplify one or more sequences comprising the nucleic acid sequence variation. The optically encoded particles may be delivered to the discrete volumes randomly resulting in a random combination of optically encoded particles in each well, or a unique combination of optically encoded particles may be specifically assigned to each discrete volume. The observable combination of optically encoded particles may then be used to identify each discrete volume.