Construction and package of recombinant adeno-associated virus encoding shRNA-Snail / 实用口腔医学杂志
Journal of Practical Stomatology
;
(6): 833-837, 2017.
Article
in Chinese
| WPRIM
| ID: wpr-697436
ABSTRACT
Objective:
To construct an adeno-associated vector expressing Snail "tough decoy" (TUD).Methods:
The nucleotide sequence of Snail gene were obtained from GenBank,then 2 cDNAs were designed and synthesized coding expression of small hairpin RNAs for Snail genes.The pAAV-U6-shRNA-Snail-TUD-EGFP expression vector was constructed by molecular biological techniques.Then pAAV-U6-shRNA-Snail-TUD-EGFP was co-transfected with Helper Free adeno-associated virus system pAAV-RC and pAAV-Helper into AAV-293 cell line to form rAAY-U6-shRNA-Snail-TUD(shRNA-Snail).The silencing effect of shRNA-Snail in Tca8113 and Cal-27 cells was detected by RT-PCR.The titer of the virus was measured.The expression level of green fluorescent protein in AAV-293 cells was monitored by the fluorescent microscopy.Results:
The result of gene sequencing showed that shRNA-Snail was successful constructed in pAAV-U6-shRNA-Snail-TUD-EGFP vector.The titer of the recombined virus was 4.4 × 1010/ml.The Snail mRNA and protein expression level was significantly reduced in Tca8113 and Cal-27 cells by rAAY-U6-shRNA-Snail-TUD-EGFP transfection.Conclusion:
rAAY-U6-shRNA-Snail-TUD-EGFP may inhibite Snail gene expression in cells.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Journal of Practical Stomatology
Year:
2017
Type:
Article
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