Electroporation-mediated nucleic acid delivery during non-embryonic stages for gene-function analysis in Anopheles sinensis

2021 ◽  
Vol 128 ◽  
pp. 103500
Author(s):  
Lin-Rong Che ◽  
Zheng-Bo He ◽  
Yan Liu ◽  
Zhen-Tian Yan ◽  
Bao-Zhu Han ◽  
...  
Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Decai Tuo ◽  
Peng Zhou ◽  
Pu Yan ◽  
Hongguang Cui ◽  
Yang Liu ◽  
...  

Abstract Background Cassava is an important crop for food security and industry in the least-developed and developing countries. The completion of the cassava genome sequence and identification of large numbers of candidate genes by next-generation sequencing provide extensive resources for cassava molecular breeding and increase the need for rapid and efficient gene function analysis systems in cassava. Several plant virus-induced gene silencing (VIGS) systems have been developed as reverse genetic tools for rapid gene function analysis in cassava. However, these VIGS vectors could cause severe viral symptoms or inefficient gene silencing. Results In this study, we constructed agroinfection-compatible infectious cDNA clones of cassava common mosaic virus isolate CM (CsCMV-CM, genus Potexvirus, family Alphaflexiviridae) that causes systemic infection with mild symptoms in cassava. CsCMV-CM was then modified to a viral vector carrying the Nimble cloning frame, which facilitates the rapid and high-throughput cloning of silencing fragments into the viral genome. The CsCMV-based vector successfully silenced phytoene desaturase (PDS) and magnesium chelatase subunit I (ChlI) in different cassava varieties and Nicotiana benthamiana. The silencing of the ChlI gene could persist for more than two months. Conclusions This CsCMV-based VIGS system provides a new tool for rapid and efficient gene function studies in cassava.


2013 ◽  
Vol 8 (8) ◽  
pp. 1551-1566 ◽  
Author(s):  
Huaiyu Mi ◽  
Anushya Muruganujan ◽  
John T Casagrande ◽  
Paul D Thomas

2004 ◽  
Vol 12 (4) ◽  
pp. 157-161 ◽  
Author(s):  
Vincent M. Bruno ◽  
Aaron P. Mitchell

Insects ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 12
Author(s):  
Chenchen Lu ◽  
Zhiqing Li ◽  
Li Chang ◽  
Zhaoming Dong ◽  
Pengchao Guo ◽  
...  

Polyamidoamine (PAMAM) dendrimers are emerging as intriguing nanovectors for nucleic acid delivery because of their unique well-defined architecture and high binding capacity, which have been broadly applied in DNA- and RNA-based therapeutics. The low-cost and high-efficiency of PAMAM dendrimers relative to traditional liposomal transfection reagents also promote their application in gene function analysis. In this study, we first investigated the potential use of a PAMAM system in the silkworm model insect. We determined the binding property of G5-PAMAM using dsRNA and DNA in vitro, and substantially achieved the delivery of dsRNA and DNA from culture medium to both silkworm BmN and BmE cells, thus leading to efficient knockdown and expression of target genes. Under treatments with different concentrations of G5-PAMAM, we evaluated its cellular cytotoxicity on silkworm cells, and the results show that G5-PAMAM had no obvious toxicity to cells. The presence of serum in the culture medium did not affect the delivery performance of DNA and dsRNA by G5-PAMAM, revealing its convenient use for various purposes. In conclusion, our data demonstrate that the PAMAM system provides a promising strategy for delivering dsRNA and DNA in cultured silkworm cells and promote its further application in individuals.


Plant Methods ◽  
2011 ◽  
Vol 7 (1) ◽  
pp. 20 ◽  
Author(s):  
Wilma Sabetta ◽  
Vittorio Alba ◽  
Antonio Blanco ◽  
Cinzia Montemurro

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