Nanoparticle-based antisense delivery systems for transient gene silencing in plants and microalgae

Overview

This solution addresses the need for cost-effective gene silencing technologies to improve plant tolerance to abiotic stress conditions such as salt stress and nitrogen deprivation. The approach uses nanoparticle-based delivery systems to transport antisense oligonucleotides into plant cells, enabling transient silencing of target genes that naturally suppress growth under stress. By targeting transcription factors from Myb and Tubby-like protein families identified through integrative proteomics and metabolomics analysis, the technology aims to enhance cellular growth performance in both microalgae and land plants. The platform leverages microalgae as a translational model for agricultural applications, offering a scalable pathway to improve crop resilience.

Technical specifications

Key features:

• Two nanoparticle types for antisense delivery: gold nanoparticles and chitosan nanoparticles
• Antisense oligonucleotides tagged with Cy5 for tracking and validation
• Target gene families include Myb and Tubby-like transcription factors with homologues in land plants
• Transformation methods optimized for nanoparticle internalization in microalgae cells
• Gene silencing effectiveness validated by qRT-PCR
• Target identification through high-resolution mass spectrometry, nuclear proteomics, and machine learning-based integrative analysis
• DNA cis-element motif detection for predicting transcription factor target genes

How it works:

The system delivers antisense molecules via nanoparticles into cells, where they bind to complementary mRNA sequences to transiently silence target gene expression. Nanoparticle internalization methods and antisense oligo release kinetics are optimized to maximize silencing efficiency. Candidate genes were selected based on proteomic and metabolomic profiling under photoautotrophic and mixotrophic conditions.

Technology readiness level

The technology is at an early-to-mid stage of development. Initial validation has been completed through successful microalgae transformation using gold nanoparticles. Proteomics, nuclear proteomics, and metabolomics datasets have been generated under controlled abiotic stress conditions with statistical significance (p<0.01). Future validation steps include synthesizing both gold and chitosan nanoparticle variants attached to antisense oligos, testing internalization methods in microalgae, optimizing antisense release, and conducting proof-of-concept gene silencing experiments on two candidate genes to demonstrate improved cell growth performance. Translation to land plants represents the next development milestone.