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Gene Synthesis

Gobind Khorana and coworkers, more than 30 years ago, in 1972 synthesized the first complete gene, a yeast tRNA. Also, the laboratories of Herbert Boyer and Alexander Markham performed the first synthesis of the peptide- and protein-coding genes. Since then, DNA synthesis methods have become a dominating technic in modern molecular biology, playing a pivotal role in synthetic biology.

Since 2007 synthetic biologists have demonstrated that not only can partial or whole chromosomes of bacteria be transplanted between bacterial cells, but scientists could also successfully synthesize a whole bacterial chromosome.

In March 2014, research scientists in Jef Boeke's group in the Langone Medical Centre at New York University achieved synthesizing one of the S. cerevisiae 16 yeast chromosomes. The research group named this chromosome III synIII.

How was this possible?

The experimental strategy used involved replacing the genes in the original chromosome with synthetic gene versions and finally integrating the finished human-made chromosome into a yeast cell. However, the design and creation of 273,871 base pairs of DNA, which contained fewer base pairs than the 316,667 pairs in the original chromosome, was required. These examples illustrate the power of existing gene synthesis techniques enabling the assembly of large DNA and RNA sequences from chemical synthetic oligonucleotide building blocks. Current oligonucleotide synthesis and enzymatic gene assembly methods and techniques are now optimized to synthesize small operons, plasmids, and viruses from scratch with relative ease. Hughes et al. in 2011 argued that these techniques are quickly becoming a cornerstone of modern molecular and synthetic biology methods.


The modern automated production of chemically synthesized DNA oligonucleotides or oligos, gene fragments, or gene blocks with high accuracy at various lengths and scales now allows the synthesis of whole genes or genomes. The field of synthetic biology ushered in an era enabling the creation of new functional genes, genetic networks, and entire genomes from genetic building blocks or artificial gene fragment now available from Biosynthesis via custom gene synthesis.

After Watson and Crick's fundamental paper in 1953 described the basic structure and chemical nature of DNA, the primary form of the double helix contains two single strands running antiparallel to each other. The two DNA strands have nucleotide chains containing sugar, phosphate, and heterocycles as part of gene building blocks. In recent years, synthetic biology developments created economic and reliable options for designing and synthesizing genes, operons, and eventually complete genomes.

However, for research in synthetic biology to succeed, accurate, economical, and high-throughput gene and genome synthesis products are essential to allow for the development of new 
tools useful to synthetic biology and biotechnology. The enzymatic assembly of chemically synthesized overlapping oligonucleotides that span the entire length of the gene construct enables synthesizing large gene blocks or whole genes.

This process needs millions of high-quality synthetic DNA oligonucleotides to construct artificial biological systems. A few decades ago, oligonucleotide synthesis was considered a rather exotic part of biochemical science pursued by only a few scientific experts. However, custom gene synthesis has now become an integral part of the arsenal of molecular biological technics. The last decade has seen developments in DNA or gene synthesis automation which can now produce more than 1000 base pairs in length.

Together with modern methods of gene isolation, sequencing, and expression, gene synthesis is playing a significant part in the advances achieved in gene technology. De novo gene synthesis is now considered the key enabling technology for synthetic biology; however, it requires the availability of genetic elements that do not exist in nature.


Biosynthesis offers custom gene synthesis to produce tailor-made synthetic oligonucleotides, gene fragments, and genes that permit the synthesis of altered or even novel proteins when using a de novo protein design and modern gene-technological methods. 


Bio-Synthesis provides high-quality gene synthesis services based on an optimized technology platform, producing double-stranded synthetic genes. All synthetic fragments are validated to ensure 100% sequence accuracy.

Applications of synthetic genes

1. Optimize codon usage in a gene or cDNA for enhanced protein expression in various expression systems.

2. Substitute for commonly used cloning methods, such as PCR.

3. Cloning of genes or cDNAs that are difficult to obtain.

4. Synthesize cDNA fragment quantities necessary for the production of custom microarray gene chips.

5. Clone humanized mouse antibodies or recombinant single-chain antibodies.

6. Design genes for gene therapy vectors and DNA vaccines.

7. Engineer genes containing specific mutations, alternative splice variants, single nucleotide polymorphisms, and others.



Reference

Annaluru, Narayana; et al. (March 27, 2014). "Total Synthesis of a Functional Designer Eukaryotic Chromosome". Science. Vol. 344, Issue 6179, pp. 55-58. [Science https://science.sciencemag.org/content/344/6179/55 ]

Edge MD, Green AR, Heathcliffe GR et al. (August 1981). "Total synthesis of a human leukocyte interferon gene". Nature 292 (5825): 756-62. [Pubmed https://pubmed.ncbi.nlm.nih.gov/6167861/  ]

Khorana HG, Agarwal KL, Büchi H et al. (December 1972). "Studies on polynucleotides. 103. Total synthesis of the structural gene for an alanine transfer ribonucleic acid from yeast". J. Mol. Biol. 72 (2): 209-217. [Pubmed https://pubmed.ncbi.nlm.nih.gov/4571075/ ]


Hughes RA, Miklos AE, Ellington AD.; Gene synthesis: methods and applications. Methods Enzymol. 2011;498:277-309. [Pubmed https://pubmed.ncbi.nlm.nih.gov/21601682/ ]


Itakura K, Hirose T, Crea R et al. (December 1977). "Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin"  Science 198 (4321): 1056-1063. [ PubMed https://pubmed.ncbi.nlm.nih.gov/412251/ ]

Shukman, David (27 March 2014). "Scientists hail synthetic chromosome advance". BBC News. Retrieved 2014-03-28. https://www.bbc.com/news/science-environment-26768445

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