CRISPR-Enabled Plant Protoplast Platform for Rapid Protein Variant Screening
A plant-protoplast-based screening platform that enables rapid construction and parallel evaluation of recombinant protein variants. By combining molecular cloning, transient expression, and molecular/biochemical assays, the platform accelerates identification of high-performing variants before committing to stable plant transformation, reducing development timelines for plant biotechnology applications.
Overview
This platform uses plant protoplasts—plant cells with their cell walls removed—as a rapid, plant-relevant system for screening recombinant protein variants. Researchers can assemble DNA variants through molecular cloning, introduce them into protoplasts for transient expression, and evaluate performance using molecular and biochemical assays. The goal is to shorten the time needed to identify promising variants before more resource-intensive stable transformation is undertaken.
The approach is designed for parallel screening, allowing multiple variants to be tested under consistent conditions. It is particularly suited for applications in plant biotechnology where protein expression, stability, and functional activity need to be assessed quickly and reproducibly.
Technical specifications
Key features:
- DNA variant constructs are generated through molecular cloning and vector construction
- Variants are introduced into plant protoplasts for transient expression, enabling fast readouts without stable transformation
- Expression levels are evaluated using PCR and quantitative reverse transcription PCR (qRT-PCR)
- Functional performance is assessed using reporter-based assays and biochemical analysis
- Workflow is designed for parallel evaluation of multiple variants and can be progressively optimized for throughput, reproducibility, and automation
Underlying expertise: The platform builds on established capabilities in protoplast isolation and regeneration, molecular cloning, vector construction, CRISPR/Cas9 genome editing, and plant biotechnology. This foundation supports the development and optimization of a robust screening workflow.
Technology readiness level
The platform is at an early development stage. Core methods in protoplast handling, molecular cloning, and biochemical analysis are established, but the integrated screening workflow has not yet been validated end-to-end.
Future validation plans include developing scalable DNA assembly and construct generation, optimizing transient expression conditions, and benchmarking multiple variants in parallel using molecular and biochemical assays. The workflow will be refined for consistency and automation, with the most promising variants advanced to plant-based validation.
About SEJONG UNIVERSITY
Sejong University is a comprehensive private research university in Seoul, South Korea, with a practical, industry-facing culture. Corporate partners engage through collaborative labs and contract research facilitated by an industry-academic cooperation foundation that streamlines agreements and compliance. A central Seoul location puts talent next to major technology and manufacturing hubs, enabling internships, executive education, and fast site-to-lab collaboration. Research is backed by competitive national programs, including the National Research Foundation of Korea and government ministries, and a dedicated technology transfer office manages IP, licensing, and startup support.
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