Rna Rescue Assay
Rna Rescue Assay - Related Videos
03:06
Rescue of Infectious Virions from Cells Transfected with Bacterial Artificial Chromosome-Derived Synthetic RNA
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2026
Source: Yun, S. et al. Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses. J. Vis. Exp. (2015)This video demonstrates the recovery of infectious Japanese encephalitis virus from synthetic RNA transcribed from a bacterial artificial chromosome clone. It shows how cells are electroporated with the synthetic RNA, allowing the viral genome to direct the production of new virions. The resulting plaques formed in infected cell monolayers confirm...
12:22
A Parasite Rescue and Transformation Assay for Antileishmanial Screening Against Intracellular Leishmania donovani Amastigotes in THP1 Human Acute Monocytic Leukemia Cell Line
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Cited by 118 •
2012
A parasite-rescue and transformation assay with THP1 cells infected in vitro with Leishmania donovani has been optimized for anti-leishmanial drug screening. The assay involves differentiation of THP1 cells, infection with promastigotes, treatment with test drugs, controlled lysis of the infected macrophages, rescue of amastigotes, transformation to promastigotes and monitoring promastigote growth and proliferation with a fluorometric assay.
04:46
A Novel Rescue Technique for Difficult Intubation and Difficult Ventilation
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2011
We describe a technique to maintain oxygenation and ventilation using an endotracheal tube inserted nasally to the level of the naso-pharynx while sealing the mouth and nares for successful positive pressure ventilation.
04:38
A Novel Technique of Rescuing Capsulorhexis Radial Tear-out using a Cystotome
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Cited by 4 •
2011
Capsulorhexis is an important step in phacoemulsification surgery. A surgeon creates a continous curvilinear tear on the anterior lens capsule by controlling the tearing vector forces. A peripherally extended tear is a serious complication. This video demonstrates a novel technique of rescuing capsular radial tear out using a cystotome.
05:07
RNA-Protein Pull-Down Assay to Isolate RNA-Binding Proteins via Affinity Extraction
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2025
This video demonstrates an in vitro RNA pull-down assay to identify RNA-binding proteins (RBPs), which interact with the adenylate-uridylate-rich element (ARE) sequences in mRNA. The target RBPs from a cell lysate are mixed with an RNA probe to form RNA-protein complexes. The complexes are isolated via affinity purification utilizing the affinity of the desthiobiotin label of the RNA probe to a streptavidin-labeled magnetic bead.
FAQs
Successful recovery after introducing candidate messenger RNA links the disrupted gene to the observed developmental defect. If embryos or cells regain normal morphology, cell behavior, or tissue formation, the result supports a direct functional connection rather than merely showing that the gene is present. This makes rescue especially useful for interpreting loss-of-function experiments.
Mutant or engineered RNA variants can separate the functions of particular protein domains, transcript regions, or regulatory features. Researchers compare whether each variant restores development as effectively as the candidate RNA. Differences in recovery indicate that the altered feature contributes to the gene’s developmental activity, helping identify which molecular elements are required for function.
Comparisons establish whether developmental recovery is associated with the candidate RNA rather than with the experimental background. A gene-specific knockdown creates the phenotype to be tested, while untreated or other control groups provide reference outcomes. Examining morphology, cell behavior, or tissue formation across these groups strengthens interpretation of whether rescue is specific and meaningful.
Researchers first disrupt or reduce the target gene in embryos or cells, then introduce synthetic messenger RNA encoding the candidate gene, often in the same experimental system. They compare development with the knockdown condition and appropriate controls, assessing features such as morphology, cell behavior, and tissue formation. The observed recovery is then interpreted as functional rescue.
This assay is useful when researchers need to validate that a loss-of-function phenotype reflects disruption of a particular gene. Introducing the corresponding RNA tests whether restoring gene expression can recover developmental outcomes. The approach can therefore connect a gene to processes such as normal morphology, cellular behavior, or tissue formation while providing functional evidence beyond gene-disruption results alone.
By testing both a candidate messenger RNA and engineered variants, researchers can examine gene function at several levels. Complete recovery supports the candidate gene’s role, whereas incomplete or absent recovery with a modified RNA can point to essential domains, transcripts, or regulatory features. These comparisons help map the molecular requirements underlying developmental processes and refine interpretations of gene loss.