Predicting Gene Silencing Through the Spatiotemporal Control of siRNA Release from Photo-responsive Polymeric Nanocarriers

Chad T. Greco; Thomas H. Epps, III; Millicent O. Sullivan

doi:10.3791/55803

Predicting Gene Silencing Through the Spatiotemporal Control of siRNA Release from Photo-responsi...

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Predicting Gene Silencing Through the Spatiotemporal Control of siRNA Release from Photo-responsive Polymeric Nanocarriers

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11:53 min

July 21, 2017

DOI: 10.3791/55803-v

Chad T. Greco¹, Thomas H. Epps, III^1,2, Millicent O. Sullivan¹

¹Department of Chemical and Biomolecular Engineering,University of Delaware, ²Department of Materials Science and Engineering,University of Delaware

Overview

This study presents a novel method utilizing photo-responsive block copolymers to achieve efficient spatiotemporal control of gene silencing. The approach allows for predictable changes in gene expression through simple siRNA release assays and kinetic modeling.

Key Study Components

Area of Science

Gene silencing
Photo-responsive materials
Biotechnology

Background

Gene silencing is crucial for regulating protein expression.
Traditional methods may have off-target effects.
Photo-responsive polymers offer a new approach to control gene expression.
Understanding structure-function relationships can enhance delivery systems.

Purpose of Study

To predict siRNA-mediated gene silencing efficiencies.
To facilitate control over protein expression levels.
To adapt assays for various stimuli-responsive systems.

Methods Used

Preparation of siRNA solutions.
Utilization of photo-responsive block copolymers.
Conducting siRNA release assays.
Kinetic modeling to predict gene expression changes.

Main Results

Efficient spatiotemporal control of gene silencing achieved.
No detectable off-target effects observed.
Changes in gene expression can be predicted accurately.
Method adaptable to other stimuli-responsive systems.

Conclusions

This method enhances control over gene expression.
It provides a foundation for drug discovery and regenerative medicine.
Future applications may include various therapeutic strategies.

Frequently Asked Questions

What are photo-responsive block copolymers?

They are materials that respond to light stimuli, allowing for controlled release of siRNA.

How does this method improve gene silencing?

It allows for precise control over the timing and location of gene silencing without off-target effects.

Can this method be adapted for other systems?

Yes, the assays can be modified to test a variety of stimuli-responsive delivery systems.

What is the significance of kinetic modeling in this study?

Kinetic modeling helps predict changes in gene expression based on siRNA release rates.

What potential applications does this research have?

It may lead to advancements in drug discovery and regenerative medicine technologies.

We present a novel method that uses photo-responsive block copolymers for more efficient spatiotemporal control of gene silencing with no detectable off-target effects. Additionally, changes in gene expression can be predicted using straightforward siRNA release assays and simple kinetic modeling.

The overall goal of these methods is to unable the prediction of small interfering RNA mediated gene silencing efficiencies and facilitate control over the resulting protein expression levels, in a spatial temporal manner. This method can elucidate structure function relationships and stimuli responsive delivery vehicles. Gaining better control over binding versus release, can unlock translatable platforms and drug discovery, as well as regenerative medicine technologies.

The main advantages of this technique are the changes in gene expression can be controlled and predicted on the base as a simple, siRNA release assays and kinetic modeling. Though these methods employ novel photo-responsive polymers, this set of assays can be readily adapted to test the wide variety of other stimuli responsive systems. To begin, prepare a 32 micrograms per milliliter siRNA solution, by adding siRNA to a 20 micromolar HEPES solution at pH six.

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