While several inhibitors targeting RNA polymerase II (Pol II) kinases have been applied for inhibiting RNA Pol II phosphorylation, there are few approaches for inducing RNA Pol II hyperphosphorylation. Here, we present a protocol for constructing the INTS8 degradation tag (dTAG) system combined with ectopic expression of N-terminally truncated INTS8 (INTS8-ΔN) in DLD-1 cells. We describe steps for INTS8-dTAG cell line construction, validation of knockin and degradation, and INTS8-ΔN rescue.
View Article and Find Full Text PDFGene expression in metazoans is controlled by promoter-proximal pausing of RNA polymerase II, which can undergo productive elongation or promoter-proximal termination. Integrator-PP2A (INTAC) plays a crucial role in determining the fate of paused polymerases, but the underlying mechanisms remain unclear. Here, we establish a rapid degradation system to dissect the functions of INTAC RNA endonuclease and phosphatase modules.
View Article and Find Full Text PDFRecent advances, specifically in the understanding of the biomechanical properties of the cornea and its response to diseases and surgical interventions, have significantly improved the safety and surgical outcomes of corneal refractive surgery, whose popularity and demand continue to grow worldwide. However, iatrogenic keratectasia resulting from the deterioration in corneal biomechanics caused by surgical interventions, although rare, remains a global concern. On one hand, in vivo biomechanical evaluation, enabled by clinical imaging systems such as the ORA and the Corvis ST, has significantly improved the risk profiling of patients for iatrogenic keratectasia.
View Article and Find Full Text PDFSci Total Environ
December 2018
While the application of anammox-based process for mesophilic sidestream treatment is at present the state of the art and mainstream treatment at ambient temperature is also in development, the feasibility of thermophilic anammox process is still unclear. This study investigated the effects of salinity on the thermotolerance of mesophilic anammox sludge. In batch activity tests, 45 °C seems to be the critical temperature for the tolerance of mesophilic anammox consortia without acclimatization or amendments.
View Article and Find Full Text PDFAppl Microbiol Biotechnol
February 2014
In this study, the effects of both transient and long-term inorganic carbon (IC) addition on the anaerobic ammonium oxidation (ANAMMOX) process under pseudo-steady-state and substrate inhibitions were analyzed using reactor performance and measures of sludge activity. Compared with the nitrogen removal rate (NRR) of 3.42 kg N m(-3) day(-1) in the control bioreactor (ICDR) without IC, the peak NRR reached 21.
View Article and Find Full Text PDFIn this study, the recovery performance of two anaerobic ammonium oxidation (ANAMMOX) reactors (R1, R2) that were previously subjected to phenol and sulfide for nearly 200 days with respective levels of 12.5-50 and 8-40 mg L(-1) and then operated in the absence of these suppressors was investigated. High nitrogen removal rates of greater than 36 kg-Nm(-3)d(-1) were achieved through the 81 and 75 days restoration of R1 and R2, respectively.
View Article and Find Full Text PDFThis paper aims to investigate the enhancement effect of low intensity intermittent ultrasound irradiation on the efficiency of anaerobic ammonium oxidation (ANAMMOX) process at ambient temperature. With intermittently irradiated (ultrasound intensity of 0.19 w/cm(2), exposure time of 0.
View Article and Find Full Text PDFZhonghua Kou Qiang Yi Xue Za Zhi
March 2007
Objective: To investigate the inhibition of telomerase activity and cellular proliferation in tongue cancer TCCA-8113 cell lines by antisense human tankyrase-1 RNA treatment, and explore the possibility of the tankyrase-1 as a target of gene therapy for tongue cancer.
Methods: The replication deficient retrovirus expressing tankyrase-1 antisense RNA was constructed to infect the TCCA-8113 cells. Tankyrase-1 expression was examined by RT-PCR.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi
August 2005
Objective: Survivin highly overexpresses in the most of human tumors, and it may play an important role in the development of tumor. The aim of this study was to explore the effects of survivin antisense oligonucleotide (ASODN) on the proliferation and the apoptosis of human Hep-2 cell.
Methods: Hep-2 cells were transfected with survivin ASODN mediated by lipofectamine, MTT [3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide] method was used to observe the cell growth inhibitory rate, the expressions of survivin mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot respectively.