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Potential Application of TALENs against Murine Cytomegalovirus Latent Infections

1
Department of Neurosurgery, Mackay Memorial Hospital, Taipei 10449, Taiwan
2
Department of Medicine, Mackay Medicine College, New Taipei City 25245, Taiwan
3
Comparative Biochemistry program, University of California, Berkeley, CA 94720, USA
4
National Applied Research Laboratories, Taipei 10636, Taiwan
*
Author to whom correspondence should be addressed.
Viruses 2019, 11(5), 414; https://doi.org/10.3390/v11050414
Received: 22 March 2019 / Revised: 24 April 2019 / Accepted: 30 April 2019 / Published: 3 May 2019
(This article belongs to the Special Issue Antiviral Agents)
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Abstract

Cytomegalovirus (CMV) infections are still a global health problem, because the latent viruses persist in humans and cause recurring diseases. Currently, there are no therapies for CMV latent infections and the therapies for active infections are limited by side effects and other problems. It is impossible to eradicate latent viruses in animals. HCMV (human CMV) is specific to human diseases; however, it is difficult to study HCMV due to its host specificity and long life cycle. Fortunately, MCMV (murine CMV) provides an excellent animal model. Here, three specific pairs of transcription activator-like effector nuclease (TALEN) plasmids (MCMV1–2, 3–4, and 5–6) were constructed to target the MCMV M80/80.5 sequence in order to test their efficacy in blocking MCMV lytic replication in NIH3T3 cell culture. The preliminary data showed that TALEN plasmids demonstrate specific targeting and cleavage in the MCMV M80/80.5 sequence and effectively inhibit MCMV growth in cell culture when the plasmid transfection is prior to the viral infection. The most specific pairs of TALEN plasmids (MCMV3–4) were further used to confirm the negative regulation of latent MCMV replication and gene expression in Balb/c mice. The injection of specific TALEN plasmids caused significant inhibition in the copy number level of immediately early gene (ie-1) DNA in five organs of mice, when compared with the controls. The result demonstrated that TALENs potentially provide an effective strategy to remove latent MCMV in animals. View Full-Text
Keywords: cytomegalovirus; latent infection; TALEN; Surveyor nuclease mutation detection assay; ie-1 gene; quantitative real-time PCR cytomegalovirus; latent infection; TALEN; Surveyor nuclease mutation detection assay; ie-1 gene; quantitative real-time PCR
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Chen, S.-J.; Chen, Y.-C. Potential Application of TALENs against Murine Cytomegalovirus Latent Infections. Viruses 2019, 11, 414.

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