Gene Knockdown Techniques

Publication Title: 
The Biochemical Journal

The free radical theory of aging proposes that ROS (reactive oxygen species) are major driving forces of aging, and are also critically involved in cellular senescence. Besides the mitochondrial respiratory chain, alternative sources of ROS have been described that might contribute to cellular senescence. Noxs (NADPH oxidases) are well-known sources of superoxide, which contribute to the antimicrobial capabilities of macrophages, a process involving the prototypical member of the family referred to as Nox2.

Author(s): 
Lener, Barbara
Kozie?, Rafa?
Pircher, Haymo
H¸tter, Eveline
Greussing, Ruth
Herndler-Brandstetter, Dietmar
Hermann, Martin
Unterluggauer, Hermann
Jansen-D¸rr, Pidder
Publication Title: 
Nature

Activating AMPK or inactivating calcineurin slows ageing in Caenorhabditis elegans and both have been implicated as therapeutic targets for age-related pathology in mammals. However, the direct targets that mediate their effects on longevity remain unclear. In mammals, CREB-regulated transcriptional coactivators (CRTCs) are a family of cofactors involved in diverse physiological processes including energy homeostasis, cancer and endoplasmic reticulum stress.

Author(s): 
Mair, William
Morantte, Ianessa
Rodrigues, Ana P. C.
Manning, Gerard
Montminy, Marc
Shaw, Reuben J.
Dillin, Andrew
Publication Title: 
PloS One

Although caloric restriction (CR) has been shown to increase lifespan in various animal models, the mechanisms underlying this phenomenon have not yet been revealed. We developed an in vitro system to mimic CR by reducing glucose concentration in cell growth medium which excludes metabolic factors and allows assessment of the effects of CR at the cellular and molecular level.

Author(s): 
Li, Yuanyuan
Tollefsbol, Trygve O.
Publication Title: 
Aging Cell

Aging and age-related diseases can be viewed as the result of the lifelong accumulation of stress insults. The identification of mutant strains and genes that are responsive to stress and can alter longevity profiles provides new therapeutic targets for age-related diseases. Here we reported that a Drosophila strain with reduced expression of ribose-5-phosphate isomerase (rpi), EP2456, exhibits increased resistance to oxidative stress and enhanced lifespan. In addition, the strain also displays higher levels of NADPH.

Author(s): 
Wang, Ching-Tzu
Chen, Yi-Chun
Wang, Yi-Yun
Huang, Ming-Hao
Yen, Tzu-Li
Li, Hsun
Liang, Cyong-Jhih
Sang, Tzu-Kang
Ciou, Shih-Ci
Yuh, Chiou-Hwa
Wang, Chao-Yung
Brummel, Theodore J.
Wang, Horng-Dar
Publication Title: 
Oncology Reports

SIRT1 is the human orthologue of SIR2, a conserved NAD-dependent protein deacetylase that regulates longevity in yeast and in Caenorhabditis elegans. Overexpression of SIRT1 in cancer tissue, compared with normal tissue, has been demonstrated, suggesting that SIRT1 may act as a tumor promoter. The function of SIRT1 in liver cancer has not been elucidated. In the present study, SIRT1 re-expression or knockdown was induced in hepatoma cell lines and liver normal cell lines.

Author(s): 
Wang, Hanning
Liu, Hao
Chen, Kaiyun
Xiao, Jinfeng
He, Ke
Zhang, Jinqian
Xiang, Guoan
Publication Title: 
PloS One

Mutations in the fused in sarcoma/translated in liposarcoma gene (FUS/TLS, FUS) have been identified in sporadic and familial forms of amyotrophic lateral sclerosis (ALS). FUS is an RNA-binding protein that is normally localized in the nucleus, but is mislocalized to the cytoplasm in ALS, and comprises cytoplasmic inclusions in ALS-affected areas. However, it is still unknown whether the neurodegeneration that occurs in ALS is caused by the loss of FUS nuclear function, or by the gain of toxic function due to cytoplasmic FUS aggregation.

Author(s): 
Sasayama, Hiroshi
Shimamura, Mai
Tokuda, Takahiko
Azuma, Yumiko
Yoshida, Tomokatsu
Mizuno, Toshiki
Nakagawa, Masanori
Fujikake, Nobuhiro
Nagai, Yoshitaka
Yamaguchi, Masamitsu
Publication Title: 
Gene

Previous studies have shown that polyphenols might be potent neuroprotective agents in Drosophila melanogaster wild type Canton-S acutely or chronically treated with paraquat (PQ), a selective toxin for elimination of dopaminergic (DAergic) neurons by oxidative stress (OS), as model of Parkinson's disease (PD).

Author(s): 
Bonilla-Ramirez, Leonardo
Jimenez-Del-Rio, Marlene
Velez-Pardo, Carlos
Publication Title: 
Methods in Molecular Biology (Clifton, N.J.)

Nutrient availability and nutrient-dependent biochemical signals represent major determinants of cellular senescence and organismal aging. The present chapter describes simple procedures to reliably evaluate the response of cultured cell to nutrients through the longevity protein p66(SHC1) and the mTOR/S6K cascade, which might be used to study cellular senescence and its chemical modulation by pharmaceutical agents in vitro.

Author(s): 
Ranieri, Sofia Chiatamone
Pani, Giovambattista
Publication Title: 
Molecular Cancer Therapeutics

Sirtuins (SIRT1-7) are a highly conserved family of NAD(+)-dependent enzymes that control the activity of histone and nonhistone regulatory proteins. SIRT1 is purposed to promote longevity and to suppress the initiation of some cancers. Nevertheless, SIRT1 is reported to function as a tumor suppressor as well as an oncogenic protein. Our data show that compared with normal liver or surrounding tumor tissue, SIRT1 is strongly overexpressed in human hepatocellular carcinoma (HCC).

Author(s): 
Portmann, Simone
Fahrner, RenÈ
Lechleiter, Antje
Keogh, Adrian
Overney, Sarah
Laemmle, Alexander
Mikami, Kei
Montani, Matteo
Tschan, Mario P.
Candinas, Daniel
Stroka, Deborah
Publication Title: 
Nature Communications

Insulin-like growth factor 1 (IGF-1) is a critical regulator of many physiological functions, ranging from longevity to immunity. However, little is known about the role of IGF-1 in natural killer cell development and function. Here, we identify an essential role for IGF-1 in the positive regulation of human natural killer cell development and cytotoxicity. Specifically, we show that human natural killer cells have the ability to produce IGF-1 and that differential endogenous IGF-1 expression leads to disparate cytotoxicity in human primary natural killer cells.

Author(s): 
Ni, Fang
Sun, Rui
Fu, Binqing
Wang, Fuyan
Guo, Chuang
Tian, Zhigang
Wei, Haiming

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