Sirna Functional Screening
Sirna Functional Screening - Related Videos
10:43
siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells
0 Views •
Cited by 4 •
2014
Here, we describe a methodology to perform a targeted siRNA “ubiquitome” screen to identify novel ubiquitin and ubiquitin-like regulators of the HIF1A-mediated cellular response to hypoxia. This can be adapted to any biological pathway where a robust read out of reporter activity is available.
02:30
siRNA - Small Interfering RNAs
0 Views •
2020
Small interfering RNAs, or siRNAs, are short regulatory RNA molecules that can silence genes post-transcriptionally, as well as the transcriptional level in some cases. siRNAs are important for protecting cells against viral infections and silencing transposable genetic elements. In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the ATP-dependent...
03:04
siRNA Encapsulation into Exosomes Using Electroporation: An Electric Pulse Based Technique to Load siRNA into Exosomes
0 Views •
2023
This video describes an electroporation-based method for encapsulating siRNA into exosomes. The siRNA-loaded exosomes can then be used for therapeutic applications.
08:21
Time-lapse Imaging of Mitosis After siRNA Transfection
0 Views •
Cited by 8 •
2010
Here we describe a basic protocol to image and quantify the mitotic timing of live mammalian tissue culture cells after siRNA transfection.
10:33
Delivery of Therapeutic siRNA to the CNS Using Cationic and Anionic Liposomes
0 Views •
Cited by 18 •
2016
The goal of this protocol is to use cationic/anionic liposomes with a neuro-targeting peptide as a CNS delivery system to enable siRNA to cross the BBB. The optimization of a delivery system for treatments, like siRNA, would allow for more treatment options for prion and other neurodegenerative diseases.
FAQs
After researchers introduce an siRNA into cancer cells, the molecule is loaded into the RNA-induced silencing complex. This complex uses the siRNA sequence to recognize complementary messenger RNA, then promotes degradation of that message or suppresses its translation. The resulting reduction in gene expression allows investigators to associate a specific gene with changes in cancer-cell behavior.
Functional effects can be assessed through cell growth, viability, apoptosis, invasion, or drug sensitivity. Each readout reflects a different aspect of cancer-cell behavior, so comparing them helps distinguish genes that support proliferation from those involved in survival, metastatic behavior, or treatment response. These measurements connect gene-specific silencing with phenotypes relevant to tumor development and therapy.
A genetic dependency is a gene function that cancer cells rely on to maintain a measured phenotype. Screens can reveal such dependencies by identifying which reductions in gene expression alter growth, viability, apoptosis, invasion, or drug sensitivity. These findings help prioritize candidate therapeutic targets and clarify pathways that support tumor development or treatment resistance.
Researchers introduce a library containing siRNAs directed against individual genes into cancer cells. They then measure selected phenotypes, such as growth, viability, apoptosis, invasion, or drug sensitivity, and compare the resulting responses across targeted genes. Genes associated with notable changes become candidates for follow-up experiments designed to test whether the observed relationship is reproducible and biologically useful.
Growth and viability measurements indicate whether reducing a gene affects continued cell maintenance, while apoptosis measurements identify effects on programmed cell death. Invasion assays address behavior associated with cancer-cell movement, and drug-sensitivity measurements reveal whether a gene influences treatment response. Selecting among these outcomes lets researchers match the screen to a specific cancer-research question.
Follow-up experiments help confirm whether a candidate gene truly relates to the observed cancer phenotype rather than relying only on the initial screen. Confirmed candidates can support investigation of genetic dependencies, potential therapeutic targets, and biomarkers. In cancer research, this validation step also helps assess whether the gene may inform precision treatment strategies or explain treatment resistance.