Targeting Vector
Targeting Vector - Related Videos
16:04
A Tetracycline-regulated Cell Line Produces High-titer Lentiviral Vectors that Specifically Target Dendritic Cells
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Cited by 6 •
2013
Here, we use retroviral transduction and concatemeric transfection to create a cell line that can express the components of a lentiviral vector (LV) in the absence of tetracycline. This LV encodes GFP and is pseudotyped with a glycoprotein, SVGmu, which is specific for a receptor on dendritic cells.
07:00
Targeting of Deep Brain Structures with Microinjections for Delivery of Drugs, Viral Vectors, or Cell Transplants
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Cited by 23 •
2010
In this article, we show a method to make glass capillary needles with a 50-μm lumen. This technique significantly reduces the brain damage, minimizes passive diffusion of drugs and allows a precise targeting into the rodent brain.
08:22
Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference
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Cited by 2 •
2013
This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
09:20
Vectors in Multiple Directions
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2023
Source: Nicholas Timmons, Asantha Cooray, PhD, Department of Physics & Astronomy, School of Physical Sciences, University of California, Irvine, CA This experiment demonstrates how vectors add and subtract in multiple directions. The goal will be to analytically calculate the addition or subtraction of multiple vectors and then to experimentally confirm the calculations. A vector is an object with both magnitude and direction. The magnitude of a vector is simply denoted as the length,...
FAQs
Homologous arms guide the construct to its intended genomic location. During homologous recombination, matching DNA sequences align the vector with the corresponding chromosome region, allowing the payload to replace or insert sequence at that locus. This locus-specific alignment distinguishes targeted modification from introducing DNA without a defined genomic destination.
The payload determines what biological change the construct can produce. A replacement sequence can alter the target gene, whereas a reporter insertion can make gene activity or regulation analyzable through an added reporting element. A selectable marker provides a way to identify cells associated with the construct. Thus, payload choice links vector design to the experimental question.
Selecting a defined locus connects the introduced change to a particular genomic region, gene, or regulatory context. This precision helps researchers relate an observed phenotype or expression change to the intended modification rather than to an unspecified site of DNA introduction. As a result, targeting vectors support more controlled studies of gene function and regulation.
A basic workflow begins by choosing the genomic region and designing homologous arms around it. Researchers then place the intended payload, such as a replacement sequence or selectable marker, between those arms and deliver the construct into cells or a model organism. Homologous recombination can then incorporate the planned modification for subsequent biological analysis.
Targeting vectors can support several experimental designs, including gene knockout, gene replacement, reporter insertion, and conditional allele generation. These options allow investigators to remove gene function, introduce an alternative sequence, examine gene activity or regulation, or create a genetically controlled form of an allele. The selected design determines the type of biological question the system can address.
Researchers apply targeting vectors in cultured cells and model organisms to investigate development, disease mechanisms, drug responses, and potential therapeutic strategies. The resulting systems provide controlled genetic changes that can be examined in relevant biological settings. Their value comes from connecting a defined genomic modification with changes in gene function, regulation, or broader biological behavior.