Genomic Targeting
Genomic Targeting - Related Videos
08:40
Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
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Cited by 1 •
2012
The Target ID Library is a plasmid-based, genome-wide collection of cloned cDNA used to identify miRNA targets. Here we demonstrate its use and application.
11:19
Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets
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Cited by 13 •
2010
Here we are presenting a chromatin immunoprecipitation (ChIP) procedure for genome-wide location analysis of protein isoforms that differ in a histone-binding domain. We are applying it to ChIP-Seq analysis to identify the targets of the KDM5A/JARID1A/RBP2 histone demethylase.
10:44
Generating Whole Bacterial Genomes from Clinical Samples using a Target Enrichment Workflow
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Cited by 1 •
2025
Here, we present a protocol to enable whole-genome sequencing of bacterial sexually transmitted infections from clinical samples using target enrichment. This novel, syndromic panel-based method overcomes challenges of abundant human DNA and low bacterial loads, facilitating genomic surveillance for Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum, and Mycoplasma genitalium.
07:05
Chromatin Immunoprecipitation to Identify Target Protein Binding Sites on Genomic DNA
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2025
In this video, we demonstrate the chromatin immunoprecipitation technique to identify protein binding sites on specific regions of the genomic DNA of oligodendrocyte precursor cells via the selective immunoprecipitation of protein-bound chromatin fragments. This method helps to study the interaction of several regulatory proteins, including transcription factors involved in gene regulation.
09:51
Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
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Cited by 18 •
2016
Genome editing of human pluripotent stem cells (hPSCs) can be done quickly and efficiently. Presented here is a robust experimental procedure to genetically engineer hPSCs as exemplified by editing the AAVS1 safe harbor locus to express EGFP and introduce antibiotic resistance.
FAQs
Sequence recognition provides the address for the intervention. In the example described, a guide RNA pairs with a complementary DNA target and directs a nuclease to that location, where cleavage occurs. The selected sequence therefore determines which genomic region is altered, while the cell’s subsequent repair pathway helps determine whether the result is disruption, replacement, or another designed change.
The two pathways support different experimental goals after nuclease cleavage. Nonhomologous end joining is associated with gene disruption, whereas homology-directed repair supports precise sequence replacement. Choosing or relying on one repair outcome changes the type of genetic alteration produced, so repair pathway behavior is central when interpreting whether an experiment created a loss of function or an intended sequence change.
A defined genomic location lets researchers connect a planned DNA change with a measurable genetic consequence. By disrupting a gene, replacing a sequence, or inserting a reporter gene, they can examine how genotype relates to phenotype and how genetic regulation operates. This makes genomic targeting especially valuable for functional studies that require a specific alteration rather than a general change to DNA.
A typical workflow begins by selecting a defined DNA sequence and establishing sequence-specific recognition, such as a guide RNA complementary to the intended target. A nuclease is directed to that site and produces a DNA break. Cellular repair then generates the experimental outcome, including disruption, replacement, or reporter-gene insertion.
Depending on the selected design and repair outcome, researchers can pursue gene disruption, precise sequence replacement, or insertion of a reporter gene. These options support different questions: disruption can test gene function, replacement can examine a defined sequence or variant, and reporter insertion can help investigate activity associated with the targeted locus. The approach therefore accommodates both loss-of-function and controlled sequence-level studies.
In genetics, targeted changes can create models of inherited disorders or address disease-associated variants, allowing researchers to relate altered sequence to phenotype. The same precision supports controlled strategies in biotechnology, where defined genomic modifications are useful. These applications extend the method from basic gene-function studies to investigations of disease mechanisms and development of more controlled genetic technologies.