Sgrna Library
Sgrna Library - Related Videos
10:10
HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
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Cited by 3 •
2019
A CRISPR/sgRNA library has been applied to interrogating protein-coding genes.However, the feasibility of a sgRNA library to uncover the function of a CTCF boundary in gene regulation remains unexplored. Here, we describe a HOX loci specific sgRNA library to elucidate the function of CTCF boundaries in HOX loci.
16:05
Large-Scale Screens of Metagenomic Libraries
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Cited by 9 •
2007
Metagenomic libraries archive large fragments of contiguous genomic sequences from microorganisms without requiring prior cultivation. Generating a ...
02:15
Nanoblade-Based Delivery of Nucleic Acid Cargo: A Technique to Deliver Cas9-sgRNA Complex to Target Cells via Virus-Like Particles
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2025
This video demonstrates a technique for nanoblade-based delivery of Cas9-sgRNA ribonucleoprotein complex inside target cells for genome editing. Nanoblades are virus-like particles devoid of the ability to multiply and infect neighboring cells, therefore are very useful for rapid and dose-dependent transportation of bio- and nanomaterials.
20:59
Large Insert Environmental Genomic Library Production
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Cited by 18 •
2009
Construction of a fosmid library with environmental genomic DNA isolated from the vertical depth continuum of a seasonally hypoxic fjord is described. The resulting clone library is picked into 384-well plates and archived for downstream sequencing and functional screening by the application of an automated colony picking system.
11:14
Primer-Free Aptamer Selection Using A Random DNA Library
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Cited by 20 •
2010
SELEX protocols comprise multiple rounds of selection, each of which require regeneration of bound ligands, which in turn require fixed primer sequences flanking the random library regions. These fixed primer sequences can interfere with the selection process (false positives and negatives). Here we present a primer-free protocol.
FAQs
Each guide can direct a Cas nuclease only when its complementary DNA target is adjacent to a compatible protospacer-adjacent motif. This motif therefore helps determine which genomic sites are accessible to targeting. In a library screen, motif compatibility affects which genes or regulatory regions can be examined and whether guide-directed disruption or regulation can occur.
The outcome depends on how the guide is used with the Cas protein. Guide-directed activity can produce targeted gene disruption, or it can alter gene regulation without necessarily disrupting the gene itself. This distinction allows researchers to investigate whether a phenotype depends on loss of gene function or on changes in gene control.
Guide representation connects the genetic perturbations introduced into a cell population with the phenotypes observed after screening. Changes in the representation of particular guides can be linked to cellular phenotypes or survival, helping identify genes that influence the measured response. This readout supports systematic analysis of many targets in parallel rather than isolated gene testing.
Researchers can examine how guide-associated genetic changes alter immune-cell activation and then relate those phenotypes to the targeted genes. Guides associated with altered activation point to genes that control immune responses, while patterns across multiple targets can reveal connected genetic pathways. These findings provide candidates for later functional studies in immunology.
A high-level workflow includes designing guides against many genomic targets, pairing each guide with a compatible Cas nuclease, and delivering the resulting library into cells. Researchers then assess cell phenotypes or survival and compare guide representation with those outcomes. The resulting associations identify targeted genes linked to the experimental response.
In infection research, pooled screens can identify host genes that affect pathogen entry, pathogen replication, or antiviral responses. Comparing guide representation with infection-related phenotypes or cell survival helps connect individual genetic perturbations to host-pathogen interactions. The approach can therefore expose cellular factors that support or restrict infection and prioritize pathways for functional investigation.
Genes associated with immune activation, pathogen entry, replication, or antiviral responses can become candidate therapeutic targets. A screen does not by itself establish a complete mechanism, but it highlights genetic pathways and individual genes for follow-up functional studies. This prioritization helps focus research on host factors that may be relevant to controlling infection or modifying immune responses.