Related Experiment Video
Updated: Jun 16, 2026

Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)
Published on: July 10, 2019
Different in vitro toxicities of structurally similar type I ribosome-inactivating proteins (RIPs)
Ou Sha1, David Tai Wai Yew, Tzi Bun Ng
1School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China. shaou@cuhk.edu.hk
Abstract:
This study was aimed at investigating and comparing the cytotoxicities of two structurally similar type I RIPs, namely trichosanthin (TCS) and free ricin A chain (RTA). A type II RIP, namely Ricinus communis agglutinin (RCA), was also included for comparison. The three RIPs were added separately to cultures of NIH 3T3 cells. The effective doses and time courses were analyzed using cell counts. Polyclonal antibodies against TCS and RTA were produced in rabbits and purified by a protein A-Sepharose CL-4B column. The mechanisms of cell death were determined by TUNEL, immunohistochemical staining, flow cytometry, and Western blotting. The effective doses for TCS, RTA and RCA were found to be 800, 50, and 50 nM, respectively. All three RIPs induced apoptosis. In all cases, activation of caspase-3 and caspase-8, but not caspase-9, was detected. Additionally, RTA caused in vivo tissue necrosis in rabbits after intradermal administration. Hence the mechanism of cell death due to RTA intoxication may vary depending on the experimental conditions, being necrosis in vivo and apoptosis in vitro. The present findings may shed light on the apoptotic pathway induced by RIPs. RTA may be useful for studying the shift in cell death.
More Related Videos
12:05RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
Published on: August 7, 2021
13:34Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip
Published on: September 29, 2012
Related Concept Videos
Ribosome Profiling
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Inhibitors of Bacterial Protein Synthesis
Riboswitches
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Ribozymes
Ribozymes can be...
Types of RNA
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
Leaky Scanning