Evaluation of self-amplifying mRNA platform for protein expression and genetic stability : Implication for mRNA therapies
Copyright © 2023 Elsevier Inc. All rights reserved..
The consecutive launch of mRNA vaccines like mRNA-1273, BNT 162b2, and GEMCOVAC®-19 against COVID-19 has triggered the debate of long-term expression, safety, and genomic integration of the mRNA vaccine platforms. In the present study, we examined the longevity of antigenic protein expression of mRNA-614 and mRNA-S1LC based on self-amplifying mRNA (SAM) in Expi-293F™, HEK-293 T, and ARPE-19 cells. The protein expression was checked by sandwich-ELISA, FACS, luciferase activity assay, and Western blot. The transcribed antigenic mRNA was sequenced and found to be un-mutated. Additionally, no genomic integration of the reverse transcribed mRNA was observed even up to 7 days post-transfection as verified by PCR. Furthermore, we have generated high-quality 3D structures of non-structural proteins (nsPs) in silico and the genes encoding for the nsPs were cloned and expressed using the T7 system. Findings from the current study have strengthened the fact that the alphavirus-based SAM platform has the potential to become a modality in the upcoming years.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:680 |
|---|---|
| Enthalten in: |
Biochemical and biophysical research communications - 680(2023) vom: 05. Nov., Seite 108-118 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Deo, Swarda [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
And genome integration |
|---|
| Anmerkungen: |
Date Revised 08.10.2023 published: Print-Electronic Citation Status Publisher |
|---|
| doi: |
10.1016/j.bbrc.2023.09.016 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM362354170 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM362354170 | ||
| 003 | DE-627 | ||
| 005 | 20231226091049.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.bbrc.2023.09.016 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1207.xml |
| 035 | |a (DE-627)NLM362354170 | ||
| 035 | |a (NLM)37738900 | ||
| 035 | |a (PII)S0006-291X(23)01046-X | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Deo, Swarda |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Evaluation of self-amplifying mRNA platform for protein expression and genetic stability |b Implication for mRNA therapies |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Revised 08.10.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status Publisher | ||
| 520 | |a Copyright © 2023 Elsevier Inc. All rights reserved. | ||
| 520 | |a The consecutive launch of mRNA vaccines like mRNA-1273, BNT 162b2, and GEMCOVAC®-19 against COVID-19 has triggered the debate of long-term expression, safety, and genomic integration of the mRNA vaccine platforms. In the present study, we examined the longevity of antigenic protein expression of mRNA-614 and mRNA-S1LC based on self-amplifying mRNA (SAM) in Expi-293F™, HEK-293 T, and ARPE-19 cells. The protein expression was checked by sandwich-ELISA, FACS, luciferase activity assay, and Western blot. The transcribed antigenic mRNA was sequenced and found to be un-mutated. Additionally, no genomic integration of the reverse transcribed mRNA was observed even up to 7 days post-transfection as verified by PCR. Furthermore, we have generated high-quality 3D structures of non-structural proteins (nsPs) in silico and the genes encoding for the nsPs were cloned and expressed using the T7 system. Findings from the current study have strengthened the fact that the alphavirus-based SAM platform has the potential to become a modality in the upcoming years | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a And genome integration | |
| 650 | 4 | |a Non-structural proteins (nsPs) | |
| 650 | 4 | |a Self-amplifying mRNA | |
| 650 | 4 | |a mRNA vaccine | |
| 700 | 1 | |a Desai, Kaushik |e verfasserin |4 aut | |
| 700 | 1 | |a Patare, Aishwarya |e verfasserin |4 aut | |
| 700 | 1 | |a Wadapurkar, Rucha |e verfasserin |4 aut | |
| 700 | 1 | |a Rade, Saniya |e verfasserin |4 aut | |
| 700 | 1 | |a Mahudkar, Siddhi |e verfasserin |4 aut | |
| 700 | 1 | |a Sathe, Madhura |e verfasserin |4 aut | |
| 700 | 1 | |a Srivastava, Shalini |e verfasserin |4 aut | |
| 700 | 1 | |a Prasanna, Pragya |e verfasserin |4 aut | |
| 700 | 1 | |a Singh, Ajay |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Biochemical and biophysical research communications |d 1960 |g 680(2023) vom: 05. Nov., Seite 108-118 |w (DE-627)NLM000000035 |x 1090-2104 |7 nnns |
| 773 | 1 | 8 | |g volume:680 |g year:2023 |g day:05 |g month:11 |g pages:108-118 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.bbrc.2023.09.016 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 680 |j 2023 |b 05 |c 11 |h 108-118 | ||