Related Experiment Videos
Aggregation and Disaggregation of Morpholino Oligomers in Solution.
Garrick Chow1, Paul A Morcos2, Hong M Moulton3
1Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, 105 Magruder Hall, Corvallis, OR, 97331, USA.
Methods in Molecular Biology (Clifton, N.J.)
|April 2, 2017
Summary
Morpholino oligomers, used to regulate gene expression, can form soluble aggregates in water, reducing their effectiveness over time. This study identifies aggregation as sequence-, temperature-, and time-dependent and offers a method to break down these aggregates.
Area of Science:
- Molecular Biology
- Biochemistry
- Drug Development
Background:
- Morpholino oligomers are potent antisense agents for gene expression regulation.
- A Morpholino-based drug is FDA-approved for Duchenne muscular dystrophy treatment.
- Aqueous solutions of some Morpholinos exhibit decreased antisense activity over time.
Purpose of the Study:
- To investigate the hypothesis that soluble aggregate formation causes decreased Morpholino activity.
- To analyze the aggregation behavior of specific Morpholino sequences in water.
- To develop a method for detecting and reversing Morpholino aggregation.
Main Methods:
- Size exclusion chromatography was employed to analyze three Morpholino sequences.
- Morpholino solutions were incubated in water to observe time-dependent aggregation.
- Factors influencing aggregation, including sequence, temperature, and time, were assessed.
Main Results:
- Two out of three analyzed Morpholino sequences formed soluble aggregates in aqueous solutions over time.
- The extent of Morpholino aggregation was found to be dependent on the specific sequence.
- Aggregation kinetics were influenced by incubation temperature and duration.
Conclusions:
- Soluble aggregate formation is a likely cause for the observed decrease in antisense activity of some Morpholinos.
- The aggregation process is sequence-, temperature-, and time-dependent.
- A straightforward procedure was developed to detect and break down Morpholino aggregates, restoring monomeric forms.