What are Transgenic Species?

An enormous development in genetic engineering came in 1973 through transgenic species when Stanley Cohen and Herbert Boyer worked in collaboration to engineer the first genetically engineered organism (i.e. mouse) successfully.

GloFish is another instance of transgenic species that has been manipulated via recombinant DNA technology in which DNA molecules belonging to two distinct species are combined together.

Foreign DNA (i.e. transgene) is referred to as DNA from the other species, or otherwise, recombinant DNA from the alike species that is being modified in the laboratory and then reintroduced. The method of the creation of transgenic cells or organisms to become whole organisms possessing a permanent change in their germ line is known as either transformation or transfection. However, transformation also indicates the process by which mammalian cell becomes cancerous, whereas transfection also indicates the process by which DNA is introduced into cells in culture (eukaryote or bacterial) for temporary use not for germline changes).

Transgenic organisms are considered vital research tools and are most often utilised during the exploration of a gene’s function. Trans genesis is related to the medical practices of gene therapy, wherein DNA is transferred to the cells of a patient for treating a disease.

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What is a Transgenic Species?

In simpler terms, transgenic refers to the transfer of particular genes from one organism to the other through gene-splicing that needs a vector for transferring the desired gene of interest from the host to the recipient. Before moving any further into the details, you must be well aware of the exact transgenic species definition. A transgenic species can be defined as the species whose genome has been manipulated by transferring a gene or genes from the other species or breed.

The picture below demonstrates two transgenic mice placed on both sides of a plain mouse. In this picture, transgenic mice are genetically modified organisms such that they carry a green coloured fluorescent protein glowing green in the presence of blue light.

These transgenic animals are frequently utilised as models in the laboratory in biomedical research. Approximately, 95 percent of those animals are genetically manipulated rodents, predominantly mice. They serve as significant tools to carry out the research in the context of human diseases for the understanding function of genes with regards to the susceptibility of diseases, and progression, and to find out the responses to the therapeutic interventions.

Moreover, mice have been genetically modified as to generate human antibodies to be utilised as therapeutics. Between 2006 and 2011, out of eleven monoclonal antibody drugs, seven drugs with FDA approval were derived from the transgenic mice.

How are Transgenic Species Produced?

For making a transgenic cell, first of all, DNA is transferred across the membrane of the cell without causing any destruction to the cell. In a few cases, naked DNA is transferred into the cell with the addition of DNA in the medium and temporarily incrementing the membrane porosity by electroporation. In another case of larger cells, naked DNA is microinjected into a cell. Other methodologies use vectors for the transportation of DNA across the membrane. Generally, the foreign DNA is a complete unit of expression including its own cis-regulators and the gene to be transcribed.

To generate a stable transgenic eukaryote, foreign DNA has to be inserted inside the chromosomes of the host. This foreign DNA enters the nucleus of the host and recombines with one of the chromatids of the host.

To generate the multicellular organisms having all transgenic cells and stably inherited transgenes, the originally transformed cell must be a gamete or must undergo development into tissues producing gametes. These transgenic gametes are eventually mated to generate homozygous, and transgenic offspring. The transgene presence can be confirmed via PCR or Southern blotting, and its expression can be monitored through RT-PCR, western blotting, and RNA blotting.

Transgenic Plant Production

Transgenic plants are more commonly produced by Agrobacterium-mediated transformation. For instance: Agrobacterium tumifaciens is a soil bacteria that injects its own tumor-inducing plasmid (T_i) into the host plant cells. The natural plasmid T_i encodes the growth-promoting genes causing a gall (i.e. tumor) for its formation on the plant that also provides an environment for the proliferation of pathogens.

T_i plasmid can be engineered by removing the tumor-inducing genes and appending restriction sites to make the insertion of desired DNA convenient. The engineered version is known as T-DNA (transfer-DNA) plasmid. A linear fragment of such plasmid including conserved right border (RB) and left border (LB) DNA sequences, is transferred by the bacterium. After getting transported into the nucleus, linear T-DNA recombines with the host DNA.

In the case of Arabidopsis or several other species, flowers are dipped in Agrobacterium suspension, and around 1% of the resultant seeds will undergo transformation. In other plant species, hormones induce cells to form a callus to which Agrobacterium is applied leading to the transformation of a few cells.

In the case of the recalcitrant plant species, other techniques like particle bombardment is used wherein DNA is attached non-covalently to small metallic particles that are accelerated into callus tissue by compressed air. In all transformation methodologies, the presence of any selectable marker is useful to distinguish transgenic cells from non-transgenic cells.

Transgenic Mouse Production

In this case, stem cells are taken out from a mouse embryo. In its place, a transgenic DNA construct is being transferred via electroporation, and homologous recombination occurs in the nucleus.

In the figure shown below, Stem cells, taken out from the embryo, are transfected with a transgenic construct bearing a neomycin resistant gene (neo^r) lined by two DNA segments homologous to the desired gene. In the transgenic cell’s nucleus, foreign DNA undergoes recombination with the targeted gene causing disruption of the targeted gene and leading to the introduction of a selectable marker. Cells carrying neo^r will survive selection. These neo^r i.e. neomycin resistant cells are transplanted into the other embryo that grows out into a chimera within the foster mother.

Human gene therapy

In accordance with the science help experts, several ethical issues and risks are connected with germline transformation due to which somatic cell transfection is mostly focused. This technology adds new genetic material to the cells. If a manipulated gene leads to a faulty or missing protein, gene therapy can add a normal gene copy to recover the functioning of the protein. Otherwise, the therapy can append a distinct gene that gives instructions for a protein that aids in the normal functioning of the cell despite the genetic modification.

The vectors can also be injected or directly provided intravenously into a particular tissue where it is consumed by individual cells. In another case, the patient's cells can be taken out and exposed to the vector in a lab. The cells comprising the vector are returned to the patient. For the successful treatment, new genes delivered by vectors make a functioning protein and will correct errors of DNA and restore protein functioning.

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References

Pinkert, C. A. (2014). Transgenic animal technology: a laboratory handbook. Newnes.

Sarkar, C., Jamaddar, S., Zulfiquar, T. N., & Mondal, M. (2020). Boon and Bane of Transgenic Animal: A Brief Review. Eur. J. Med. Health Sci, 2(2), 21-27.Tatiana, M. (2021). New naturally transgenic plants: 2020 update. Biological Communications, 66(1), 36-46.