Construction of lentiviral shRNA expression vector targeting phospholipase D2 (PLD2) gene

2011 ◽  
Vol 10 (66) ◽  
Author(s):  
Lian
Keyword(s):  
Expression Vector ◽  
Shrna Expression ◽  
Shrna Expression Vector ◽  
Phospholipase D2 ◽  
Vector Targeting

Induced Apoptosis Of Multidrug Resistance Leukemia K562/A02 Cells By Magnetic Nanoparticle Of Fe3O4 Copolymerizating JNK1 shRNA Expression Vector and Daunorubicin

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4925-4925
Author(s):  
Wei Zhang ◽  
Baoan Chen ◽  
Xiangfeng He ◽  
Rong Fu
Keyword(s):  
Multidrug Resistance ◽  
Protein Level ◽  
Magnetic Nanoparticle ◽  
Expression Vector ◽  
Caspase 3 ◽  
Chromatin Condensation ◽  
Dapi Staining ◽  
Shrna Expression ◽  
Chemotherapeutic Regimen ◽  
Shrna Expression Vector

Abstract Currently, serious toxicity and poor antitumor efficacy resulting from the induction of multidrug resistance (MDR) in hematological malignancies and solid tumors are common hindrances to successful systemic chemotherapy. One of the important mechanisms of tumorigenesis and resistance to anticancer drugs was blockade of the apoptosis-inducing pathway. In order to overcome tumor MDR while maintaining good antitumor activity, biomaterial combine with gene therapy provides a novel strategy for the treatment of MDR.The aim of this study was to explore the induction of apoptosis by magnetic nanoparticle of Fe3O4 (MNPs-Fe3O4), a novel delivery system for chemotherapeutic regimen that is chemically stable, environmentally friendly, and noncytotoxic, copolymerizating JNK1shRNA expression vector with DNR, which could overcome siRNA technique instability and enhance DNR chemotherapeutic efficiency and provide a new field to reverse tumor MDR. JNK1shRNA expression vector was constructed and screened the most effective plasmid and transfected the cells. Typical apoptotic characteristics and apoptosis effect of MNPs-Fe3O4 and PGM-3 with DNR were investigated by DAPI staining and FCM assay, respectively. Additionally, drug accumulation and apoptosis relative-genes were evaluated in K562/AO2 leukemia cells.The results showed that PGCsilencer-U6-neo-GFP-GV102 shRNA/JNK1 was successfully constructed, which was confirmed by sequencing that expression of JNK1 gene in mRNA and protein level that had the less in PGM-3 than that of PGM-1 or PGM-2. Meanwhile, MNPs-Fe3O4 and PGM-3 with DNR (DNR/PGM-3/MNPs-Fe3O4) could synergistically induced typical apoptotic characteristics of chromatin condensation and apoptotic body with DAPI staining, and a higher rate of apoptosis were detected in K562/A02 cells treated with DNR/PGM-3/MNPs-Fe3O4. Intracellular DNR was higher in K562/A02 cells treated with DNR/PGM-3/MNPs-Fe3O4 than that of DNR/MNPs-Fe3O4 by FCM analysis.  Further study demonstrated that both DNR/PGM-3/MNPs-Fe3O4 and DNR/PGM-3 reduced the gene transcriptions and protein expressions of bcl-2, enhanced that of bax and caspase-3, decreased survivin in protein level, and has no different change in mRNA level by qRT-PCR and Western blotting analyses, respectively. These findings showed that the combination of MNPs-Fe3O4 with JNK1 shRNA expression vector and DNR could induce apoptosis of K562/A02 cells through elevating the ratio of bax/bcl-2, activating caspase-3, and decreasing survivin in protein level. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Construction of an All-in-one Double-conditional shRNA Expression Vector

2019 ◽  
Vol 31 (1) ◽  
pp. 41-50
Author(s):  
Yoshiko KUDO ◽  
Hideto OYAMADA ◽  
Tomoyuki MATSUOKA ◽  
Ayami INAGAKI ◽  
Hiromichi TSUCHIYA ◽  
...  
Keyword(s):  
Expression Vector ◽  
Shrna Expression ◽  
Shrna Expression Vector

scholarly journals Development of a species-specific RNA polymerase I-based shRNA expression vector

2006 ◽  
Vol 35 (2) ◽  
pp. e10-e10 ◽  
Author(s):  
M. S. Brenz Verca ◽  
P. Weber ◽  
C. Mayer ◽  
C. Graf ◽  
D. Refojo ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Expression Vector ◽  
Rna Polymerase I ◽  
Shrna Expression ◽  
Shrna Expression Vector ◽  
Species Specific ◽  
Polymerase I
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