Aav Microinjection
Aav Microinjection - Related Videos
08:12
Zebrafish Microinjection Techniques
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2023
One of the major advantages to working with zebrafish (Danio rerio) is that their genetics can be easily manipulated by microinjection of early stage embryos. Using this technique, solutions containing genetic material or knockdown constructs are delivered into the blastomeres: the embryonic cells sitting atop the yolk of the newly fertilized egg. Delivery into the cytoplasm is achieved either through direct injection into the blastomere, or via natural cytoplasmic movements that occur after a...
12:10
Microinjection of Xenopus Laevis Oocytes
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Cited by 14 •
2009
Here we demonstrate cytoplasmic microinjection of Xenopus laevis oocytes with a nuclear import substrate, as well as preparation of the injected oocytes for visualization by thin-sectioning electron microscopy.
10:06
Personalized Needles for Microinjections in the Rodent Brain
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Cited by 3 •
2018
We describe here a protocol for microinjection in the rodent brain that uses quartz needles. These needles do not produce detectable tissue damage and ensure reliable delivery even in deep regions. Moreover, they can be adapted to research needs by personalized designs and can be re-used.
02:24
AAV-Mediated Gene Delivery for Visualization of Compromised Retinal Membrane Integrity
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2026
Source: Vacca, O., et al. Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease. J. Vis. Exp. (2015).This video demonstrates the method of AAV-mediated gene delivery in a transgenic mouse retina, followed by tissue dissection and staining to assess inner limiting membrane integrity.
17:39
Microinjection of A. aegypti Embryos to Obtain Transgenic Mosquitoes
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Cited by 56 •
2007
In this video, Nijole Jasinskiene demonstrates the methodology employed to generate transgenic Aedes aegypti mosquitoes, which are vectors for dengue fever. The techniques for correctly preparing microinjection needles, dessicating embryos, and performing microinjection are demonstrated.
FAQs
Cell-type-specific promoters restrict expression of the engineered gene to the intended cellular population, rather than relying only on where the injection is placed. This adds a molecular layer of selectivity to the anatomical targeting provided by stereotaxic coordinates. In neuroscience, that combination helps researchers examine or manipulate particular components of a neural circuit with greater precision.
After entering target cells, engineered AAV genes can produce fluorescent labels, tracing tools, calcium or voltage sensors, or proteins that alter neural activity. The selected payload therefore determines what the experiment can reveal: cell location or connectivity, activity dynamics, or consequences of activating and controlling neural populations. This makes vector design central to the study question.
Long-lasting access allows labeled or genetically modified neural populations to remain available for examining brain function, connectivity, and disease mechanisms. It also supports experiments in which researchers need to observe activity or manipulate a circuit after gene expression has been established. Thus, the value of the method extends beyond the initial injection to sustained circuit-level investigation.
The workflow begins by selecting stereotaxic coordinates for the brain region of interest. A fine needle then delivers a small volume of the chosen recombinant AAV vector into that location. The vector enters cells in the targeted region and supports expression of its engineered gene. Researchers can then use the resulting labeling, sensing, tracing, or activity-manipulation capability.
Researchers should match the engineered gene and promoter to the desired readout or intervention. Fluorescent constructs support labeling, tracing tools address connectivity, and calcium or voltage sensors report neural activity. If the goal is to alter activity, optogenetic or chemogenetic proteins provide the relevant control strategy. This alignment links the biological question to the molecular payload delivered by the injection.
It is especially useful when a study requires access to a defined brain region and selected cell populations at the same time. The approach can connect anatomical targeting with measurements of activity, mapping of connectivity, or controlled changes in neural activity. These capabilities make it relevant to investigations of brain function, circuit organization, and disease mechanisms.
By placing an engineered gene in a selected brain region and cell population, researchers can examine how specific neural circuits relate to disease mechanisms. Fluorescent labeling and tracing can reveal relevant organization, while activity sensors or activity-manipulation proteins can test circuit function. The resulting regional and cellular access helps link neural changes with broader brain processes.