Targeted Dna Insertion
Targeted Dna Insertion - Related Videos
08:38
Targeted DNA Methylation Analysis by Next-generation Sequencing
0 Views •
Cited by 83 •
2015
Bisulfite amplicon sequencing (BSAS) is a method for quantifying cytosine methylation in targeted genomic regions of interest. This method uses bisulfite conversion paired with PCR amplification of target regions prior to next-generation sequencing to produce absolute quantitation of DNA methylation at a base-specific level.
02:55
Immunoprecipitation of Target Protein-DNA Complexes from Motor Neuron Lysate
0 Views •
2025
This video demonstrates the immunoprecipitation of DNA fragments crosslinked with target protein from stem cell-derived motor neuron lysates using target protein-specific antibody-coated magnetic beads.
07:05
Chromatin Immunoprecipitation to Identify Target Protein Binding Sites on Genomic DNA
0 Views •
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.
15:28
Primer Extension Capture: Targeted Sequence Retrieval from Heavily Degraded DNA Sources
0 Views •
Cited by 41 •
2009
We present a method of targeted ancient DNA sequence retrieval, which we used to reconstruct the complete mitochondrial genomes of five Neandertal individuals. Comparison of these sequences with present day humans suggests that Neandertals had a long term low effective population size.
04:24
TALEN-Mediated Targeted Gene Integration: Using Engineered Sequence-Specific Nucleases for the Precise Insertion of Fluorescent Protein Gene into hiPSCs
0 Views •
2025
This video describes a precise genome editing technique in human induced pluripotent stem cells, or hiPSCs, using TALENs to create double-stranded breaks at a targeted locus, inducing homology-directed repair for fluorescence protein gene integration.
FAQs
A programmable nuclease, such as a CRISPR-associated enzyme, recognizes the selected genomic site and creates a break there. That break provides the local entry point for repair, allowing the introduced sequence to be incorporated at the intended locus rather than positioned without regard to genomic coordinates. This targeting step is central to controlling the genetic change.
The donor DNA template supplies the sequence researchers want to add and can guide repair by homology-directed recombination. In this process, matching sequence information helps connect the donor with the broken target region. The template therefore determines what is inserted, while the nuclease determines where the repair event is initiated, linking sequence design with positional control.
Compared with random insertion, this approach fixes the genomic position under investigation, giving researchers greater control over where the engineered sequence resides. That positional control helps connect an observed phenotype to the intended genetic alteration, making results easier to interpret. Random insertion provides less certainty about how genomic location contributed to the outcome.
A conceptual workflow starts by choosing a genomic site and preparing a programmable nuclease for that target. Researchers also provide a donor DNA template carrying the desired sequence. The nuclease-generated break then engages a repair route, such as homology-directed recombination or another insertion pathway, to produce the planned genomic change.
Targeted DNA insertion can support gene tagging, correction of disease-associated variants, transgene integration, and functional analysis of regulatory elements. These applications use positional control for different purposes, including modifying a specified variant, placing a chosen transgene, or examining how selected regulatory DNA contributes to gene function and regulation.
The donor template may guide homology-directed recombination, but the process can also use another insertion pathway. This distinction matters because the repair route is part of how the chosen sequence becomes incorporated after nuclease cutting. Considering that route helps researchers interpret how an engineered locus was produced in gene-function or regulatory-element studies.