Sniper-screen System
Sniper-screen System - Related Videos
09:44
High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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2015
Here we describe a cell-based reporter gene assay as a valuable tool to screen chemical libraries for compounds modulating post-transcriptional control mechanisms exerted through 3’ UTR.
08:50
Genetic Screens
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2023
Genetic screens are critical tools for defining gene function and understanding gene interactions. Screens typically involve mutating genes and then assessing the affected organisms for phenotypes of interest. The process can be “forward”, where mutations are generated randomly to identify unknown genes responsible for the phenotypes, or it can be “reverse”, where specific genes are targeted for mutation to observe what phenotypes are produced.Here, JoVE reviews various types of genetic...
08:10
Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
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Cited by 7 •
2016
This work presents a method of high-throughput screening using a universal genetic enzyme screening system that can be theoretically applied to over 200 enzymes. Here, the single screening system identifies three different enzymes (lipase, cellulase, and alkaline phosphatase) by simply changing the substrate used (p-nitrophenyl acetate, p-nitrophenyl-β-D-cellobioside, and phenyl phosphate).
09:40
RNAi Screening to Identify Postembryonic Phenotypes in C. elegans
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Cited by 9 •
2012
We describe a sensitized method to identify postembryonic regulators of protein expression and localization in C. elegans using an RNAi-based genomic screen and an integrated transgene that expresses a functional, fluorescently tagged protein.
11:37
Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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Cited by 21 •
2019
Here, we present a protocol to optimize CRISPR-Cas9 to achieve a higher specificity without the loss of on-target activity. We use a directed evolution approach called Sniper-screen to find a mutant Cas9 with the desired characteristics. Sniper-Cas9 is compatible with truncated single-guide RNAs and delivery in a ribonucleoprotein format, well-known strategies for achieving higher specificities.
FAQs
Targeted perturbations focus the screen on selected genes or variants, allowing researchers to examine specific functional possibilities. Systematic perturbations broaden the search across a wider genetic space and can reveal candidates that were not selected in advance. This distinction affects the scope of the experiment and the type of genotype–phenotype relationships the analysis can uncover.
The phenotype provides the measurable readout that connects a genetic perturbation with a biological outcome. Selection identifies perturbations associated with a chosen outcome, while measurement can capture differences in phenotype across experiments. A clearly defined readout therefore determines which DNA changes become informative and supports comparison among candidate genes or variants.
Sequence analysis links the observed phenotype back to the DNA changes associated with the screened material. Researchers use that sequence information to identify candidate genes or variants for further interpretation. This step transforms a biological observation into genetic evidence that can support functional gene discovery and analysis of relationships between molecular changes and phenotypes.
A typical workflow begins by applying targeted or systematic genetic perturbations, followed by selecting or measuring the resulting phenotypes. Researchers then analyze associated sequence changes and connect those changes with the observed outcomes. Organizing the experiment around these stages helps maintain a clear path from perturbation, through phenotype assessment, to candidate gene or variant identification.
Researchers can apply the Sniper-screen System when they need to determine how genetic changes influence a measurable biological phenotype. It is relevant to functional gene discovery, where candidate genes are linked to outcomes, and to variant analysis, where DNA changes are examined in relation to their effects. The same framework supports larger-scale genetic investigations.
By associating molecular alterations with observable phenotypes, the system can help researchers investigate pathways underlying development or disease. Candidate genes and variants identified through screening provide points for examining how genetic changes relate to these biological processes. This makes the approach useful for connecting large-scale genetic data with pathway-level questions in genetics research.