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Methods and Applications of Gene Synthesis

HomeEducationalMethods and Applications of Gene Synthesis

Gene synthesis is a procedure that refers to the base-by-base synthesis of a DNA strand. It is different from DNA replication, which occurs through Polymerase Chain Reaction in the cells. Rather than using a template strand, gene synthesis depends on a single-stranded molecule that functions as a template onto which nucleotides are added systematically. Gene synthesis has become a fundamental technology in the field of synthetic biology. Nowadays, you can easily order synthetic genes online through a gene synthesis service.

Gene Synthesis Methods

There are numerous methods used in gene synthesis, and they all vary depending on the complexity and length of the DNA as well as other non-technical factors such as automation capability and intellectual property rights.

However, all the currently available technologies used in the synthesis of genes depend on the chemical synthesis of oligonucleotides, which is the first step for generating the building blocks required for enzymatic assembly. Oligonucleotides are commercially produced and sold since the process of making them is automated, negating the need for individuals to make their own.

1. PCR-Based Reactions

Polymerase Chain Reaction is the most appealing method of gene synthesis. Its foundation lies on the DNA polymerase property of creating a new strand of DNA that corresponds to the provided template strand. DNA polymerase requires a primer since it can add nucleotide only onto an established 3′-OH group. As a result, the researcher can target specific sections of the DNA, such as a particular gene, to synthesize. The particular sequence of interest can be produced in billions using PCR. However, PCR-based methods are error-prone as a result of mutations introduced during oligonucleotide synthesis and sequence. Nevertheless, PCR techniques such as sequential chain reaction (SCR) are still the most viable commercially.

2. Slonomics Technology

Slonomics is a novel gene synthesis technique that relies on universal double-stranded DNA as the building blocks. Industrial robotic machines are used to assemble the desired gene using a highly-standardized biochemical process. Slonomics is considered to be superior to other gene synthesis techniques because it utilizes a computerized and highly standardized process in addition to quality raw materials. It eliminates many challenges to gene synthesis and is efficient and reliable even for genes that are difficult to synthesize.

3. Phosphoramidite Method

The phosphoramidite method uses a 5’ hydroxy terminus to which nucleotides are added sequentially to form an oligonucleotide chain. The procedure involves de-protection, coupling, capping, and oxidation steps. Without capping, more than one nucleotide would be added to the sequence. The cap allows the process to be systematic and, when removed, the process can refresh and continue. This method is, however, not ideal for long oligonucleotides as errors increase with the length of the oligonucleotide.

4. Chip-Based Technology

Building DNA on a computer chip is a new method that promises to revolutionize the gene synthesis industry. Other methods rely on synthesizing genes in individual wells then assembling them into strands. The chip-based synthesis relies on an electrochemical, heat-controlled technology that allows for the accommodation of several independent temperature control sites, which can link with thermally labile reagents to recreate the oligonucleotide synthesis processes of the phosphoramidite cycle with high selectivity. This method also enables easy detection of errors, which can be rectified during assembly. The result is the synthesis of higher fidelity double-stranded DNA.

There are other methods available for gene synthesis, including the Fok I method, ligation of predefined reusable duplexes of phosphorylated overlapping oligos, and a modified ligase chain reaction method. The downside of these procedures is that they cannot produce the best results unless you use phosphorylated, polyacrylamide gel (PAGE) purified oligos. The process for developing these oligonucleotides is time-consuming, labor-intensive, and costly, making these methods unsuitable for commercial use.

Applications Of Gene Synthesis

Gene synthesis plays a critical role in modern molecular biology, particularly synthetic biology. Scientists can generate new genetic pathways, whole genes, and even entire genomes.

1. Bioengineering

Synthetic biology is an emerging science that deals with the design, development, and improvement of biological systems to make these processes easier, reliable, and more predictable. Bioengineering uses engineering principles to manipulate biological systems to develop or create bio-based materials, therapeutics, chemicals, and additional sources of energy and food. Bioengineering is particularly useful in medical and biomedical fields where gene synthesis is used in the development of new medical devices and medical techniques. The ability to sequence genomes has led to the creation of artificial living systems. Of interest is the engineering of parallel metabolic systems that allow scientists to program cells that can interfere with the natural chemical pathways. This has enabled the production of drugs and other chemicals cost-effectively.

2. Neuroscience

In neuroscience, gene synthesis is mainly used for research studies. With the help of codon-optimized channelrhodopsins, it is possible to use optogenetic techniques to visualize neuronal pathways. This way, gene variants developed by gene synthesis can be used to study various pathways, such as the neuronal signaling mechanisms that control learning behaviors and synapse plasticity. In this area, gene synthesis has played a vital role in enabling the understanding of brain structures, diversity of proteins, and even human behavior.

gene

3. Vaccine Design

Gene synthesis has become a cornerstone of vaccine research. It is applied in basic immunological and microbiological studies that play an important role in vaccine design and even development. These studies have given hope in developing novel vaccines that can combat persistent and emerging viruses such as seasonal Influenza A and Ebola Virus, respectively. In cancer studies, these vaccines are promising in developing drugs that can boost the immune system against oncogenic viruses.

4. Therapeutic Antibodies

Gene synthesis is used in the production of antibody-based drugs, which have shown promises in the treatment of many diseases, including autoimmune and inflammatory disease, and cancer. Monoclonal antibodies, which are therapeutic antibodies, can repress, activate, or change endogenous immune reactions to certain molecules or cells. Drugs are developed based on these interactions.

Conclusion

Gene synthesis is now an easy process thanks to the advancement of technology. Better technologies have been developed for synthesizing genes. Similarly, as scientists gain more knowledge in this area, more applications are discovered. However, gene synthesis remains to be most useful in the medical field.

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Jessica S
Jessica S
Jessica Smith has been writing articles for e-business and e-lance sites for more than 4 years. Her educational background is Masters in English and journalism which gives her a broad platform to write on a variety of topics with ease and efficiency. She is an independent writer especially enjoys writing on fashion, lifestyle, health, and medical niche.

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