Related Experiment Video
Updated: Aug 10, 2026
11:53
Predicting Gene Silencing Through the Spatiotemporal Control of siRNA Release from Photo-responsive Polymeric Nanocarriers
Published on: July 21, 2017
Gene expression control by temperature with thermo-responsive polymeric gene carriers
M Kurisawa1, M Yokoyama, T Okano
1Institute of Biomedical Engineering, Tokyo Women's Medical University, Kawada-cho 8-1, Shinjuku-ku, Tokyo 162-8666, Japan.
Summary
This study developed a thermo-responsive copolymer for gene delivery. Lowering incubation temperature enhanced DNA complex stability and significantly improved gene transfection efficiency in cells.
Area of Science:
- Biomaterials Science
- Polymer Chemistry
- Gene Therapy
Background:
- Gene therapy holds promise but requires efficient and safe delivery vectors.
- Developing stimuli-responsive polymers can enhance gene delivery control.
- Thermo-responsive polymers offer tunable properties based on temperature changes.
Purpose of the Study:
- To synthesize and characterize a thermo-responsive copolymer for gene delivery.
- To evaluate the effect of temperature on copolymer-DNA complex stability.
- To assess the in vitro gene transfection efficiency of the copolymer at different temperatures.
Main Methods:
- Synthesis of poly(N-isopropylacrylamide-co-2-(dimethylamino)ethyl methacrylate-co-butylmethacrylate) copolymer.
- Determination of copolymer's lower critical solution temperature (LCST).
- Gel electrophoresis to analyze copolymer-plasmid DNA complex stability at varying temperatures.
- In vitro gene transfection assay in COS-1 cells using pCMV-lacZ plasmid.
Main Results:
- The synthesized copolymer (P(IP-8DA-11BM)) exhibited an LCST of 21°C, remaining soluble below this temperature and insoluble above.
- DNA was stably retained in the complex at 37°C but partially dissociated at 20°C.
- Gene transfection efficiency was significantly higher when using P(IP-8DA-11BM) at 20°C (3h) and 37°C (45h) compared to 37°C (48h).
Conclusions:
- Temperature significantly modulates the formation and dissociation of the copolymer-DNA complexes.
- The thermo-responsive copolymer P(IP-8DA-11BM) demonstrates potential as a gene delivery vector with temperature-controlled efficiency.
- Optimizing incubation temperature is crucial for maximizing gene transfection efficacy with this novel copolymer.
More Related Videos
Related Concept Videos
01:42What is Gene Expression?
Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
02:24Regulation of Expression Occurs at Multiple Steps
Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
02:33Combinatorial Gene Control
Combinatorial gene control is the synergistic action of several transcriptional factors to regulate the expression of a single gene. The absence of one or more of these factors may lead to a significant difference in the level of gene expression or repression.
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
The expression of more than 30,000 genes is controlled by approximately 2000-3000 transcription factors. This is possible because a single transcription factor can recognize more than one regulatory sequence. The specificity in gene...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
01:36What is Gene Expression?
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then processed and...
01:23Regulation of Expression at Multiple Steps
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...