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Applied Sciences
Volume 10
Issue 22
10.3390/app10228004
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Article
Peer-Review Record

Encapsulation of mRNA into Artificial Viral Capsids via Hybridization of a β-Annulus-dT20 Conjugate and the Poly(A) Tail of mRNA

Appl. Sci. 2020, 10(22), 8004; https://doi.org/10.3390/app10228004
by Yoko Nakamura 1, Yuki Sato 1, Hiroshi Inaba 1,2, Takashi Iwasaki 3 and Kazunori Matsuura 1,2,*
Reviewer 1: Anonymous
Reviewer 2: Anonymous
Appl. Sci. 2020, 10(22), 8004; https://doi.org/10.3390/app10228004
Submission received: 18 September 2020 / Revised: 9 November 2020 / Accepted: 9 November 2020 / Published: 12 November 2020
(This article belongs to the Special Issue Nucleic Acids Conjugates for Biotechnological Applications)

Round 1

Reviewer 1 Report

The aim of this work was to construct the viral artificial capsids for an mRNA delivery. This is a continuation of previous work on b-annulus peptide, however this manuscript presents interesting new data. Surprising is that: when poly(A) and dT20-SS-annulus were mixed at equimolar base concentrations, capsid formation was insufficient. Charge repulsion should not be an issue, because this is similar to unfunctionalized DNA which hybridize easily, especially in NaCl containing PBS buffer.

 

Major and minor comments:

 

Figure 4A Surprising is also the lack of EMSA shift when mRNA is missed with dT20-SS-annulus peptide mixture (line 4-7) as compared to RNA + dT20. Such bulky hydrophilic group is expected to change the gel mobility.

 

The EMSA picture of mRNA + dT20-SS-annulus peptide in one figure 4A line 4 (approximate size 500bp) is different from the one presented in figure 6B line 4 (approximate size 200bp)

 

Minor comments:

 

What indicator was used for EMSA visualization?

via is Latin and should be italic throughout the document

Author Response

Please see the attachment.

Author Response File: Author Response.docx

Reviewer 2 Report

This manuscript described the design of artificial capsids encapsulating mRNA via hybridization for a delivery carrier.  The concept (material design) is interesting in spite of little properties for mRNA delivery carrier in vitro.  The manuscript can be accepted after the following revision.

 

1) In case of mRNA in this study, because of mRNA is unstable as compared to DNA, the controlled release of encapsulated mRNA by reductive cleavage of disulfide bonds should be examined.  For example, EMSA should be carried out in the presence of reducing agent such as DTT.

 

2) If the controlled release of mRNA occurs, the time-course of the transfection experiment should be examined.   

 

3) Figure 6: Please identify lane No. in Figure 6.

Author Response

Please see the attachment.

Author Response File: Author Response.docx

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