Microinjection

Microinjection is the use of a glass micropipette to inject a liquid substance at a microscopic or borderline macroscopic level. The target is often a living cell but may also include intercellular space.

It can be used to deliver antibody targeted to a specific protein domain in order to analyze the requirement of the protein for specific cell functions such as cell cycle progression, transcription of specific genes, or intracellular transport.

It is the process of transferring genetic materials into a living cell using glass micropipettes or metal microinjection needles. Glass micropipettes can be of various sizes with tip diameters ranging from 0.1 to 10 µm. DNA or RNA is injected directly into the cell's nucleus.

Microinjection is a technique in which recombinant DNA is directly injected into the nucleus of an animal. In this, through a glass micropipette, foreign DNA is delivered directly into a living cell, oocyte or embryos of animal.

 It is a technique of delivering foreign DNA into a living cell (a cell, egg, oocyte, embryos of animals) through a glass micropipette. One end of a glass micropipette is heated until the glass becomes somewhat liquified. It is quickly stretched which forms a very fine tip at the heated end.

Bacterial and fungal cells are too small for microinjection, making it an impractical procedure.

DNA microinjection is the dominating technique leading to random integration of a transgene via the introduction of DNA into the pronucleus of a developing zygote. Following fertilization of a mouse egg, the male and female pronuclei remain separated for a few hours before they fuse to make the zygotic nucleus, thus allowing for microinjection of the desired genes into the larger male pronucleus. Eggs that can survive the injections are transferred into the oviducts of a foster mother i.e., pseudopregnant female mice for the generation of Founder mouse, from which permanent transgenic lines can be established. The presence of transgenes in the offsprings can be identified by PCR analysis or Southern blot hybridization.

Microinjection of DNA is performed shortly after fertilization. The pronuclei of mouse eggs are readily visualized using Differential Interference Contrast (DIC) optics (see image to right).  Fertilized eggs have two pronuclei, one from each gamete. The injected DNA can integrate into chromosomes in the injected pronucleus during chromosome replication.  After the DNA in each pronucleus has duplicated, the pronuclear envelopes break down, the duplicated chromosomes align on the metaphase plate and the fertilized oocyte divides to form a two-cell stage mouse embryo.

Rubin and Spradling (1982) for the first time introduced Drosophila gene for xanthine dehydrogenase into a P-element (parental element) which was microinjected with an intact helper P-element into embryo deficient for this gene. Such embryos later on developed flies with rosy colored eyes than mosaic eyes as in the first parental generation.

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Journal of  Biochemistry and Biotechnology