Guide Rna Screening
Guide Rna Screening - Related Videos
09:18
TRUE Gene Silencing: Screening of a Heptamer-type Small Guide RNA Library for Potential Cancer Therapeutic Agents
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Cited by 7 •
2016
Here we present a protocol for screening of a heptamer-type sgRNA library for potential therapeutic drugs against blood cancers.
09:19
RNA Catalyst as a Reporter for Screening Drugs against RNA Editing in Trypanosomes
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Cited by 1 •
2014
A highly sensitive ribozyme-based assay, applicable to high-throughput screening of chemicals targeting the unique process of RNA editing in trypanosomatid pathogens, is described in this paper. Inhibitors can be used as tools for hypothesis-driven analysis of the RNA editing process and ultimately as therapeutics.
02:12
Co-Infiltration Assay for Screening Pathogen Effectors that Suppress Plant RNA Silencing
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2026
Source: Shi, J., et al. Screening and Identification of RNA Silencing Suppressors from Secreted Effectors of Plant Pathogens. J. Vis. Exp. (2020)This video demonstrates a co-infiltration assay for identifying pathogen effector proteins that suppress RNA silencing in Nicotiana benthamiana 16c. A fluorescent reporter and a candidate effector gene are delivered into plant leaves via Agrobacterium. Normally, RNA silencing degrades reporter transcripts, reducing fluorescence. Suppressor effectors...
11:34
High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses
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Cited by 14 •
2014
In vitro assays to measure virus replication have been greatly improved by the development of recombinant RNA viruses expressing luciferase or other enzymes capable of bioluminescence. Here we detail a high-throughput screening pipeline that combines such recombinant strains of measles and chikungunya viruses to isolate broad-spectrum antivirals from chemical libraries.
06:48
CRISPR Guide RNA Cloning for Mammalian Systems
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Cited by 16 •
2018
Here, a simple, efficient, and cost-effective method of sgRNA cloning is outlined.
FAQs
A guide RNA provides the targeting component, directing a Cas nuclease or transcriptional effector to a selected DNA sequence. The resulting perturbation can then be linked to a cellular measurement, such as guide abundance, gene expression, survival, or another phenotype. This connection lets a screen associate particular genes or regulatory elements with observed cellular behavior.
Comparing guide representation before and after selection reveals which perturbations become more or less represented under the tested condition. In pooled screens, sequencing supplies this comparison at the library level. Changes in abundance therefore serve as a population-based readout, helping connect targeted genetic perturbations with effects on cell survival or another selected phenotype.
The choice determines the type of perturbation applied to the targeted sequence. A Cas nuclease is one possible effector, whereas a transcriptional effector provides another way to alter gene-related activity. This flexibility allows screening designs to examine targeted effects across genes or regulatory elements while matching the perturbation system to the biological question.
A typical workflow starts with a guide RNA library designed against selected DNA sequences. Researchers apply the library-based perturbations to cells, define a readout such as guide abundance, gene expression, survival, or another measurable trait, and then compare guide representation before and after selection, commonly using sequencing. The comparison identifies perturbations associated with the phenotype.
Useful interpretation depends on the chosen readout and how it changes after selection. Guide abundance can indicate differential representation, while gene expression, cell survival, or another measurable trait can report a cellular phenotype. Reading these signals together with the targeted gene or regulatory element helps identify perturbations linked to the outcome under study.
In bioengineering, the approach supports pathway optimization by revealing genetic changes associated with desired cellular properties. It also contributes to functional genomics, disease modeling, and drug-target identification. Because screens connect perturbations with measurable outcomes, they can inform the design of engineered cells with improved properties and help prioritize genes or regulatory elements for further study.