Journal
ACS OMEGA
Volume 4, Issue 1, Pages 458-464Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsomega.8b02865
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Funding
- KAKENHI from Japan Ministry of Education, Culture, Sports, Science and Technology (MEXT) [17K05611]
- Grants-in-Aid for Scientific Research [17K05611] Funding Source: KAKEN
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Encapsulation of nucleic acids is an important technology in gene delivery, construction of artificial cells, genome protection, and other fields. However, although there have been a number of protocols reported for encapsulation of short or oligomeric DNAs, encapsulation of genome-sized DNA containing hundreds of kilobase pairs is challenging because the length of such DNA is much longer compared to the size of a typical microcapsule. Here, we report a protocol for encapsulation of a ca. 60 mu m contour length DNA into several micrometer-sized polyelectrolyte capsules. The encapsulation was carried out by (1) compaction of T4 DNA with multivalent cations, (2) entrapment of DNA condensates into micrometer-sized CaCO3 beads, (3) assembly of polyelectrolyte multilayers on a bead surface, and (4) dissolution of beads resulting in DNA unfolding and release. Fluorescence microscopy was used to monitor the process of long DNA encapsulation at the level of single-DNA molecules. The differences between long and short DNA encapsulation processes and morphologies of products are discussed.
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