1.Inactivation of gene by small interference RNA
Journal of Medical Research 2003;21(1):89-93
The role of suppression of small RNA molecules in the management of malignity and viral infection on human was studies and discussed. SiRNA (small interference RNA) suppressing gene expression was described. In the year 2001, Ribopharma AG researchers had first demontrated the function of RNAi in mammal cells. SIRPLEX is appropriate with target gene, for using in the treatment of suppression of pathological gene in various genera, including human.
RNA
;
Genes
;
Gene Silencing
2.RNA interference in functional genomics and medical research.
Zili YOU ; Jian HUANG ; Ye ZHENG ; Dezhong YAO
Journal of Biomedical Engineering 2004;21(5):848-851
RNA interference (RNAi) is a post-transcriptional gene silencing process by targeting mRNA for degradation in a sequence-specific manner. This powerful platform has enormous potential in functional genomics and medical research. As a tool to knock out expression of specific genes in a variety of organisms, RNAi was used to investigate gene function in a high throughput fashion. Highly conserved in evolution RNAi appears to have evolved as a cellular defense mechanism in plants and animals to suppress viral infection, transposon jumping and endogenous aberrant genes. Exploiting the natural mechanism, the researchers can shut down disease-causing genes and develop novel therapeutics against infection, tumor and other disease.
Gene Expression Regulation
;
Gene Silencing
;
Genomics
;
RNA Interference
;
RNA, Small Interfering
;
RNA-Induced Silencing Complex
3.The mechanism and application of posttranscriptional gene silencing.
Yang LIU ; Yan JIANG ; Dai-Rong QIAO ; Yi CAO
Chinese Journal of Biotechnology 2002;18(2):140-143
The purpose of this review is to confirm the reason resulted the gene silence and explore the countermeasure avoiding the gene silence in transgene plant. The method is to divide the gene silencing into transcriptional gene silencing(TGS) and posttranscriptional gene silencing(PTGS). Several models resulted PTGS were analyzed by RNA threshold model, ectopic pairing and aberrant RNA model and ds-RNA model. The results showed that it was important to decide the phenomena of restraining transgene silencing and the mechanism of PTGS. The strategies of identification of gene function and prevention of virus were presented by RNAi and gene silencing respectively, etc.
Animals
;
Gene Silencing
;
Models, Genetic
;
RNA
;
RNA Processing, Post-Transcriptional
4.Establish a screening system for selection of mRNA target sites for HBsAg to construct siRNA with shRNA.
Zheng-Gang YANG ; Zhi CHEN ; Ning XU ; Qin NI ; Xiu-Cheng PAN ; Han-Ying JIN ; Min-Wei LI
Chinese Journal of Hepatology 2004;12(9):515-518
OBJECTIVETo find some effective short interfering RNA's sites targeting HBV surface gene sequence using shRNA expression vectors.
METHODSFour shRNA expression vectors targeting HBV surface gene sequence were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with HBs-EGFP fusion gene plasmid. The changes of HBs-EGFP image were detected by FACS and microscopy. The HBs-EGFP mRNA expression was evaluated by RT-PCR.
RESULTSFour shRNA expression vectors and HBs-EGFP fusion gene plasmid were successfully constructed. pAVU6 + 4sh579 vector inhibited the HBs-EGFP expression by 69.8% in AD293 and suppressed the HBs-EGFP mRNA expression by 74.6%.
CONCLUSIONSThe results showed that the 579 site of HBV surface gene sequence was an effective target and pAVU6 + 4sh579 vector could suppress the HBs-EGFP expression in AD293 cells
Gene Expression Regulation, Viral ; Gene Silencing ; Gene Targeting ; methods ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; RNA-Induced Silencing Complex ; genetics
5.Effects of AKR1B10 gene silence on the growth and gene expression of HCC cell line MHCC97H.
Wei WEI ; Hong-jie LIANG ; Jie-feng CUI ; Kun GUO ; Xiao-nan KANG ; Ji CAO ; Jian-jia SU ; Yuan LI ; Yin-kun LIU
Chinese Journal of Hepatology 2010;18(9):666-671
OBJECTIVETo explore the biological function and possible underlying mechanism of aldo-keto reductase family 1 member B10 (AKR1B10) gene during hepatocarcinogenesis.
METHODSA pair of chemically synthesized small interfering RNA (siRNA) targeting on AKR1B10 was transfected into liver cancer cell line MHCC97H by LipofectamineTM 2000. After confirming the interfering effects of AKR1B10-siRNAs through Quant SYBR Green polymerase chain reaction (Real-time PCR), Western blot and enzymatic activity assay, the capabilities of proliferation and apoptosis of the transfected cells were observed by CCK-8 assay and flow cytometry analysis, and the expressions of a group of tumor-related gene such as c-myc, c-fos, N-ras were observed through Real-time PCR.
RESULTSThe expressions of AKR1B10 and the enzymatic activity were down-regulated significantly in AKR1B10-siRNA-transfected cells. Compared with mock and blank control groups, cell growth in AKR1B10-siRNA-transfected group was inhibited by 26.6%+/-3.1% at 72h after transfection. The ratio of apoptotic cells was 37.3%+/-1.0% in AKR1B10-siRNA-transfected group, which was significantly higher than that in mock and blank control groups (P < 0.01). Real-time PCR showed that the expressions of oncogene c-myc, c-fos and N-ras, and the proliferation-associated gene ki-67 were down-regulated in AKR1B10-siRNA-transfected cells, while the expressions of apoptosis-promoting gene caspas-3 and bax were up-regulated.
CONCLUSIONSAKR1B10 might promote proliferation, inhibit apoptosis and then induce malignant transformation of hepatocytes by regulating the expression level of some tumor-related genes.
Aldehyde Reductase ; genetics ; Cell Line, Tumor ; Gene Expression ; Gene Silencing ; Humans ; RNA, Small Interfering ; genetics
6.Application of virus-induced gene silencing technology to investigate the phytochrome metabolism mechanism: a review.
Duo PAN ; Songyue ZHANG ; Fangyi LIU ; Qingyin TIAN ; Xiulian YANG ; Lianggui WANG ; Yuanzheng YUE
Chinese Journal of Biotechnology 2023;39(7):2579-2599
Color is an important indicator for evaluating the ornamental traits of horticultural plants, and plant pigments is a key factor affecting the color phenotype of plants. Plant pigments and their metabolites play important roles in color formation of ornamental organs, regulation of plant growth and development, and response to adversity stress. It has therefore became a hot topic in the field of plant research. Virus-induced gene silencing (VIGS) is a vital genomics tool that specifically reduces host endogenous gene expression utilizing plant homology-dependent defense mechanisms. In addition, VIGS enables characterization of gene function by rapidly inducing the gene-silencing phenotypes in plants. It provides an efficient and feasible alternative for verifying gene function in plant species lacking genetic transformation systems. This paper reviews the current status of the application of VIGS technology in the biosynthesis, degradation and regulatory mechanisms of plant pigments. Moreover, this review discusses the potential and future prospects of VIGS technology in exploring the regulatory mechanisms of plant pigments, with the aim to further our understandings of the metabolic processes and regulatory mechanisms of different plant pigments as well as improving plant color traits.
Plant Viruses/genetics*
;
Plants/genetics*
;
Gene Silencing
;
Plant Development
;
Gene Expression Regulation, Plant
;
Genetic Vectors
8.Cavin-2 Functions as a Suppressive Regulator in TNF-induced Mesenchymal Stromal Cell Inflammation and Angiogenic Phenotypes.
Bayader ANNABI ; Alain ZGHEIB ; Borhane ANNABI
International Journal of Stem Cells 2017;10(1):103-113
Tumour necrosis factor (TNF)-α activation of mesenchymal stromal cells (MSC) enhances their tumour-suppressive properties and tumour-homing ability. The molecular actors involved are unknown. We found that TNF induced MSC migration and tubulogenesis which correlated with a dose-dependent increase in Cavin-1 and Cavin-3 transcript levels. TNF triggered cyclooxygenase (COX)-2 expression, whereas specific siRNA-mediated gene silencing of Cavin-2 resulted in an amplified COX-2 expression, tubulogenesis, and migratory response partially due to a rapid and sustained increase in NF-κB phosphorylation status. Our results highlight a suppressive role for the caveolar component Cavin-2 in the angiogenic and inflammatory regulation of TNF-activated MSC.
Gene Silencing
;
Inflammation*
;
Mesenchymal Stromal Cells*
;
Necrosis
;
Phenotype*
;
Phosphorylation
;
Prostaglandin-Endoperoxide Synthases
9.Methylation Status of CpG Island of p16 in Benign, Atypical and Malignant Meningiomas.
Jeong Hoon KIM ; Jae Sung AHN ; Sang Ryong JEON ; Yong Hee SHIM ; Chang Jin KIM ; Jung Kyo LEE
Journal of Korean Neurosurgical Society 2003;33(2):126-131
OBJECTIVE: Hypermethylation of p16, a tumor suppressor gene, has been frequently detected in a variety of cancer cells and is known to represent the level of p16 transcription. In human meningiomas, genetic alterations of p16 have shown to be infrequent. The purpose of this study is to investigate the role of p16 associated with the progression of meningiomas. METHODS: Sixty-eight meningiomas(randomly sampled 29 benign, 16 atypical and 23 malignant formalin-fixed, paraffin-embedded tissues) were analyzed. We examined the molecular mechanism of inactivation of p16 in these benign, atypical and malignant meningiomas by detecting the methylation status of p16 using methylation-specific polymerase chain reaction. RESULTS: One out of 29(3.4%) revealed hypermethylation of p16 in benign meningiomas. Atypical and malignant meningiomas showed hypermethylation of p16 in 2 out of 16 cases(12.5%) and in 5 out of 23 cases(21.7%), respectively. Immunohistochemical analysis of methylation-positive tumors demonstrated that tumor cells had reduced immunoreactivity compared to normal lymphocytes. CONCLUSIONS: Our results suggest that inactivation of p16 gene plays a role in the pathogenesis of meningioma and hypermethylation is one of the processes for gene inactivation.
CpG Islands*
;
Gene Silencing
;
Genes, p16
;
Genes, Tumor Suppressor
;
Humans
;
Lymphocytes
;
Meningioma*
;
Methylation*
;
Polymerase Chain Reaction
10.Abnormalities of beta-Catenin Gene in Pilomatricomas.
Hyun Jeong LEE ; Seog Jun HA ; Jung Soo KIM ; Seung Dong LEE ; Jin Wou KIM
Korean Journal of Dermatology 2002;40(1):14-18
BACKGROUND: beta-Catenin muations were reported to play a causal role in the development of pilomatricomas. In a recent study in Caucasians, 75% of pilomatricomas had beta-Catenin mutations. OBJECTIVE: We investigated the causal role of beta-Catenin gene mutations in pilomatricomas of Koreans. METHODS: This study included 20 formalin-fixed, paraffin-embedded pilomatricomas in Koreans. Basophilic nucleated tumor cells were microdissected and, as normal controls, infiltrating inflammatory lymphocytes were microdissected from the same histologic specimens. Sequencing analysis of exon 3 of the beta-Catenin gene (CTNNB1) was performed. Immunostaining for beta-Catenin and Lef-1 was performed by the avidin-biotin-peroxidase method. RESULTS: Sequencing analysis found missense mutations (S37Y, S37C, S33C, S33F, and S37F) in CTNNB1 in 6 samples (30%) of 20 pilomatricomas. All pilomatricomas revealed intense expression of nuclear Lef-1 and nuclear and cytoplasmic beta-Catenin. CONCLUSION: Frequencies of beta-Catenin mutations were lower in our study compared with the results in Caucasians. The immunohistochemical results suggest the abnormalities in Wnt-wingless pathway resulting in stabilization or constitutive expression of beta-Catenin, but the absence of CTNNB1 mutations in 70% of our cases suggests adenomatous polyposis coli gene inactivation, or the involvement of other components of the Wnt-wingless pathway.
Adenomatous Polyposis Coli
;
Basophils
;
beta Catenin*
;
Cytoplasm
;
Exons
;
Gene Silencing
;
Lymphocytes
;
Mutation, Missense
;
Pilomatrixoma*