Development of a high-throughput platform for screening lipid nanoparticles for mRNA delivery

Nanoscale ◽  
2022 ◽  
Author(s):  
Lili Cui ◽  
Sara Pereira ◽  
Silvia Sonzini ◽  
Sally Van Pelt ◽  
Steven Romanelli ◽  
...  

mRNA lipid nanoparticles (LNPs) are at the forefront of nucleic acid intracellular delivery, as exemplified by the recent emergency approval of two mRNA LNP-based COVID-19 vaccines. The success of an...

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 615
Author(s):  
Allen Wing-Ho Chu ◽  
Cyril Chik-Yan Yip ◽  
Wan-Mui Chan ◽  
Anthony Chin-Ki Ng ◽  
Dream Lok-Sze Chan ◽  
...  

SARS-CoV-2 RT-PCR with pooled specimens has been implemented during the COVID-19 pandemic as a cost- and manpower-saving strategy for large-scale testing. However, there is a paucity of data on the efficiency of different nucleic acid extraction platforms on pooled specimens. This study compared a novel automated high-throughput liquid-based RNA extraction (LRE) platform (PHASIFYTM) with a widely used magnetic bead-based total nucleic acid extraction (MBTE) platform (NucliSENS® easyMAG®). A total of 60 pools of nasopharyngeal swab and 60 pools of posterior oropharyngeal saliva specimens, each consisting of 1 SARS-CoV-2 positive and 9 SARS-CoV-2 negative specimens, were included for the comparison. Real-time RT-PCR targeting the SARS-CoV-2 RdRp/Hel gene was performed, and GAPDH RT-PCR was used to detect RT-PCR inhibitors. No significant differences were observed in the Ct values and overall RT-PCR positive rates between LRE and MBTE platforms (92.5% (111/120] vs 90% (108/120]), but there was a slightly higher positive rate for LRE (88.3% (53/60]) than MBTE (81.7% (49/60]) among pooled saliva. The automated LRE method is comparable to a standard MBTE method for the detection of SAR-CoV-2 in pooled specimens, providing a suitable alternative automated extraction platform. Furthermore, LRE may be better suited for pooled saliva specimens due to more efficient removal of RT-PCR inhibitors.


2017 ◽  
Vol 15 (1) ◽  
pp. 53-62 ◽  
Author(s):  
Zongxiang Jia ◽  
Yan Gong ◽  
Yufang Pi ◽  
Xueying Liu ◽  
Lipeng Gao ◽  
...  

Plant Methods ◽  
2010 ◽  
Vol 6 (1) ◽  
pp. 3 ◽  
Author(s):  
Yellamaraju Sreelakshmi ◽  
Soni Gupta ◽  
Reddaiah Bodanapu ◽  
Vineeta Chauhan ◽  
Mickey Hanjabam ◽  
...  

2021 ◽  
pp. 2101391
Author(s):  
Zuzana Hejdankova ◽  
Vaclav Vanek ◽  
Frantisek Sedlak ◽  
Jan Prochazka ◽  
Audrey Diederichs ◽  
...  

2019 ◽  
Vol 47 (18) ◽  
pp. e107-e107 ◽  
Author(s):  
Daehyung Kim ◽  
Fahad Rashid ◽  
Yeonmo Cho ◽  
Manal S Zaher ◽  
I I Hwan Cho ◽  
...  

Abstract Real-time visualization of single-proteins or -complexes on nucleic acid substrates is an essential tool for characterizing nucleic acid binding proteins. Here, we present a novel surface-condition independent and high-throughput single-molecule optical imaging platform called ‘DNA skybridge’. The DNA skybridge is constructed in a 3D structure with 4 μm-high thin quartz barriers in a quartz slide. Each DNA end is attached to the top of the adjacent barrier, resulting in the extension and immobilization of DNA. In this 3D structure, the bottom surface is out-of-focus when the target molecules on the DNA are imaged. Moreover, the DNA skybridge itself creates a thin Gaussian light sheet beam parallel to the immobilized DNA. This dual property allows for imaging a single probe-tagged molecule moving on DNA while effectively suppressing interference with the surface and background signals from the surface.


2013 ◽  
Vol 5 (11) ◽  
pp. 2835 ◽  
Author(s):  
C. H. van den Kieboom ◽  
T. S. Y. van Domburg ◽  
M. I. de Jonge ◽  
G. Ferwerda ◽  
P. W. M. Hermans

2000 ◽  
Vol 5 (5) ◽  
pp. 343-351 ◽  
Author(s):  
Usha Warrior ◽  
Yihong Fan ◽  
Caroline A. David ◽  
Julie A. Wilkins ◽  
Evelyn M. McKeegan ◽  
...  

To identify inhibitors of interleukin-8 (IL-8) production, a high throughput assay was developed using the Quanti-Gene™ nucleic acid quantification kit that employs branched-chain DNA (bDNA) technology to measure the mRNA directly from cells. Unlike polymerase chain reaction and other technologies that employ target amplification, the QuantiGene system uses signal amplification. To perform the assay, various molecular probes capable of hybridizing with IL-8 mRNA were designed and synthesized. A human lung epithelial cell line was treated with interleukin-la (IL-la) to stimulate the IL-8 gene expression and the mRNA was measured using the QuantiGene system. The QuantiGene assay was sensitive, flexible, and reproducible and achieved equivalent or better sensitivity than promoter-reporter assays, and eliminated the time required for constructing a promoter-reporter system. Our data show that bDNA technology has the potential to be used as a high throughput screening assay.


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