Gene synthesis has revolutionized what was once thought to be impossible in the field of genetics. Synthesis of the first gene, a yeast tRNA, was achieved by Har Gobind Khorona and coworkers in 1972. Since then this technology has had long last impacts on genetic research and the biotechnology industry. Whether it is synthesizing small genes, ranging in lengths from a few hundred base pairs in length, to large genes, up to 100kb in length, gene synthesis has made recently daunting research aspirations achievable. Gene synthesis has a simple definition, the creation of synthetic genes within a laboratory setting, but an impactful influence on many scientific fields. One field in particular, genetic engineering, is often associated with gene synthesis. The association between these two topics is so prevalent because gene synthesis is a popular mechanism to achieve the genetic engineering sought out by researchers. Genetic engineering also has a wide range of application in many different scientific fields, many relying on gene synthesis.

Gene synthesis itself has a many applications, ranging from developing more effective vaccinations to creating large synthetic gene libraries. Synbio Technologies offers the ability to take a requested genetic sequence, in text format, and create the desired physical sequence with one hundred percent accuracy. This is done with our, patent pending, Syno^®2.0 Platform. The unique feature that this platform offers is the capability of synthesizing a genetic sequence that may not be preexisting within any organism. Previously, the sequence of interest was required to be existing in order to be extracted and later cloned. The Syno^®2.0 Platform allows us to generate the user specified sequence exactly, confirmed with Sanger sequencing, with a both time and cost effective manner. This technology is the foundation for genetic engineering. Genetic engineering is defined as the alteration of an organism’s genome using biotechnology.

This is accomplished through the incorporation of a genetic sequence that is foreign to an organism into its genome. The unique connection between gene synthesis and genetic engineering is found within this step. Gene synthesis allows us to synthetically create the gene of interest, which is then incorporated into the organism’s genome. The gene of interest can have a wide range of functions, and are specific and tailor made to the interest of the researcher. Once the gene of interest is incorporated into the organism’s genome, research can be conducted to determine effects of the newly incorporated gene on the organism. This general outline has been applied to a wide range of applications in various fields of interest.

The applications of genetic engineering can be found in many fields of ranging from gene therapy, through the use of viral vectors, to agriculture. Gene synthesis and genetic engineering have been used to study the effects of a genetic knockout in a particular gene, a method that is very commonly used in genetic research. It is seen when studying the effects of gene amplification, causing an overexpression of a gene of interest. Genetic engineering is most commonly seen in agriculture through the creation of genetically modified crops to optimize the quality as well as quantity of certain crops. This process has been going on for thousands of years through selective breeding, but recent advances in technology have allowed researchers to implement groundbreaking innovations on basic crop production. These innovations help to combat environmental pressures, such as draught or blight, as well as decreasing pesticide use by genetic modification. These modifications, accomplished by genetic engineering, have a simple goal in mind: to increase the quality and quantity of these crops which are essential in everyday life. It is through genetic engineering and gene synthesis that these innovations are possible.

The main advantage that gene synthesis has when relating to genetic engineering is the accessibility and ease of synthetically creating your gene of interest. With Synbio Technology’s Syno^®Platform the process from desired sequence to genetically modified organism is easily achieved. This process starts with the input of the researcher’s requested sequence, up to 100kb in length. The requested sequences is created with one hundred percent accuracy and verified with Sanger sequencing. This step is done with a both time and accuracy efficient pipeline. The desired sequence will then be shipped to your location within as few as five business days. In addition to the time and accuracy efficiency Synbio Technologies offers an extremely cost effective approach, with prices starting at $0.25 per base pair for gene synthesis. The combination of competitive pricing, time efficiency, and accuracy have lead Synbio Technologies to be one of the leading biotechnology companies, especially within the realm gene synthesis and its relationship with genetic engineering.

Recently, gene synthesis has become an extremely popular and effective method to conduct genetic research. This rise in popularity stems from the ease of use as well as the effective results. In short, gene synthesis can be described as the creation of a synthetic genetic sequence specified by the user. Recently, the genetic sequence of interest must have been present within an organism to be extracted and later studied. Gene synthesis allows researchers to bypass this step entirely, creating the sequence of interest and converting the sequence from text format to physical copy with ease. This accessibility has caused multiple companies to participate in this groundbreaking technology within the field of genetics. As the gene synthesis industry has grown drastically, companies have competed to become the best at synthesizing genes. At Synbio Technologies we pride ourselves as being one of the leading companies within this gene synthesis industry. This pride and confidence is mainly relying upon our, patent pending, Syno® Platforms. It is the three phases of this platform that separates us from any other company in the gene synthesis industry, allowing us to serve you with a high quality product, with an efficient time frame and competitive prices.

The first platform that Synbio Technologies offers, the Syno^®1.0 Platform, is an industrialized DNA chemical synthesis platform. Essentially, this platform specializes in the formation of oligos. An oligo can be described as a short fragment of DNA or RNA molecule. These oligos have a wide range of functions in various fields of genetic research including forensics, and genetic testing. In order to accomplish the output of high quality oligos, Synbio Technologies has four steps within this mechanism. First, the DNA sequence of interest is analyzed and designed. Once this complete, CG-base oligo synthesis is conducted. This step includes industry leading and manufacturing process which allows for the highest quality oligos. In order to increase the quality, the oligos are then purified using the proprietary purification process. Finally, there is then an additional step of quality assessment using MS analysis to verify the created oligos are what the user requested. The Syno® 1.0 Platform has a wide range of applications using this four part mechanism. Some applications include generating native DNA sequences, creating de novo DNA sequences, and constructing degenerated oligo libraries and so on. All of these processes have been utilized and proven with the use of oligo synthesis. This is just the first step in the process, which has allowed Synbio Technologies to become one of the premier companies in the gene synthesis industry. The remaining two platforms are what really separate us from the competition.

The Syno^®2.0 Platform focuses on PCR based gene synthesis and is an aspect that Synbio Technologies prides itself as being one of the leaders in the biotechnology and gene synthesis industries. Synbio Technologies is capable of synthesizing a wide range of gene lengths up to and including 100kb. The process as to how these genes are synthesized are constantly verified for accuracy as well as quality. The resulting synthesized gene product is guaranteed to be one hundred percent identity to the user specified requested sequence. The resulting synthesized gene can then be applied in various types of genetic research. One major application that differs from many others is the ability to generate de novo DNA sequences. This aspect has revolutionized genetic research since the early 1970s, increasing in quality and effectiveness along the way. The creation of these de novo DNA sequences has allowed researchers to develop more effective vaccinations, generate variant libraries, and improve the features of protein. It is the basis of so many crucial fields of genetic research. At Synbio Technologies we offer the highest quality gene synthesis product with the most competitive prices in the gene synthesis industry. We pride ourselves with the accuracy of our product which we are able to provide our users.

The third and final platform, the Syno^®3.0 Platform, specializes in oligo pool synthesis. This platform is described as a chip-based next generation DNA synthesis platform. Essentially, this platform allows us to generate the highest quality DNA fragments, de novo synthesized genes, small genomes, and variant libraries. In addition to this, the high quality output of these various genetic materials is provided to the user at the lowest prices in the gene synthesis industry. The process that this platform follows is very similar to that of the Syno^®1.0 Platform with some small and necessary alterations. First the proprietary DNA sequences are designed, analyzed and optimized. Once this is complete, the chip-based oligo pool synthesis process begins. This step utilizes the, industry leading, oligo-pool assembly and manipulation process. After that, there is proprietary error correction, allowing for a quality control and improving the resulting output. Then large fragment assembly, up to 200kb, is conducted. Once this is finished, the final step is to run a quality control including next generation sequencing. This final step allows for the resulting sequence to be verified and one hundred percent accurate. Overall, the applications of this platform are similar to that of the previous Syno^®1.0 and 2.0 Platforms. The user can generate native and de novo DNA sequences, build genes, chassis, operons, pathways and small genomes, as well as generate DNA variant libraries. In addition to these features, this platform offers a unique aspect which allows the process to be scalable. This means that it will take the same amount of time to produce 20,000 genes as it would to produce 20 genes, making the Syno^®3.0 Platform more effective. These features are what make the Syno^®3.0 Platform extremely valuable to many users.

Gene synthesis is a fundamental technique in synthetic biology, allowing for the creation of unique synthetic genes that would be difficult or impossible to obtain naturally. In order to express a synthetic gene in an efficient and controlled way, expression vectors are often used to introduce the gene into a cell, after which the cell’s own gene expression system is used to synthesize the target protein.

Common Gene Synthesis Vector Types

There are three main categories of carriers that constitute vector systems in gene synthesis engineering: plasmids, bacteriophages and viruses. One of the most widely used expression vectors is that of Escherichia coli due to its ease of growth, rapid reproduction, and ease of transformation with exogenous DNA. Others include integration vectors, bacteriophage vector promotion, or shuttle vectors in S. cerevisiae.

Occasionally, a series of pGEX vectors (-1T, -2T, -3T), GST (glutathione S-trasferase) system, pEZZ18 are utilized for specific expression systems. Several variants transformed from pBluescript Ⅱ SK(+), together with pUC18, pUC19, pUC57, are also selected for certain projects.

Synbio Technologies are able to synthesize a large variety of target DNA and clone them into any vector as requested, regardless of the specifications of the project or the approach required. We guarantee a highly accurate and high-yield product at a low cost and with rapid turnaround.

Gene Synthesis Vector Design Application

Synbio Technologies provides two main approaches for amplifying DNA sequences via PCR cloning and subcloning technology. For DNA or amino acid sequences provided by customers, Synbio Technologies will synthesize error-free DNA de novo by request. Our proprietary Syno^®gene synthesis platform can perfectly achieve enzymatic assembly through limited short DNA fragments. Even genes with challenging characteristics such as repetitive sequences, complex secondary structure, and high (>80%) or low (20%) GC content as well as long polypyrimidine runs, can be synthesized correctly.

Additionally, our professional Syno^®Codon software can compute and optimize codon selection for original sequences or specific vectors. Any synthetic vector meeting our requirements can be quickly and accurately delivered at an economical price. Synbio Technologies can insert any target gene into any site of any vector, and can guarantee a high-quality, fully accurate final product for gene synthesis.

Introduction

Over the last few decades, gene synthesis and assembly technology have developed rapidly. Since the 1960s, our capability to synthesize genes has skyrocketed from less than 100bp to more than 1,000,000bp. Gene synthesis methods and applications have a profound impact in metabolic engineering, genetic network design, and vaccine design. However, existing gene synthesis methods still have their fair share of drawbacks and inconsistencies, such as low yield, high error rate, and lack of scalability both on the size of the project and the size of the gene.

Gene Synthesis Methods

Gene synthesis methods are not able to replace each other, and each occupies its own niche depending on the requirements of the project. The following is a brief overview of several common gene synthesis methods:

• Solid-phase synthesis

The traditional oligonucleotide synthesis uses a small volume of solution processed in a column full of chemicals. The oligonucleotides are synthesized by attaching nucleotide residues stepwise to the end of the chain, one-by-one. The addition of each oligonucleotide consists of four steps: de-blocking, coupling, capping, and oxidation. The integrity of the sequence and the productivity of the synthesis are hindered for products longer than 200bp, and thus this method is generally limited by DNA sequence length. The primary advantage of this method is its high accuracy, compensating for its high expense and low output.

• Chip-based DNA synthesis

As the name implies, Chip-based synthesis utilizes microarray chips utilizing a series of electrochemical techniques. Different kinds of oligonucleotides are able to be synthesized in different specific parts of the chips, called assembly subpools. Following this piecewise synthesis, gene fragments in subpools are amplified and then aggregated and assembled into the finished product. Chip-based DNA synthesis is cheaper than solid-phase synthesis and can yield a larger amount of the target gene, but its accuracy suffers in comparison.

• PCR based enzyme synthesis

PCR-based enzyme synthesis generates gene fragments through a variety of cell systems. Using the Yeast system as an example, by using different incision enzymes and label markers, different kind of genes can be added to Yeast chromosomes. Due to the nature of gene insertion the target gene could have no limit to its length as long as the chromosomes can accommodate. This method performs well in synthesizing large gene fragments, and with the help of the cell systems the accuracy of the gene sequence is guaranteed.

Gene synthesis applications

Synthetic genes have wide implications on a variety of fields, ranging from genetic circuits to metabolic improvement to many more. As our technology advances and our understanding of genetics continues to develop, scientists could modify and design genes, The scientists have already synthesized and assembled a gene fragment of over 100kbp. When inserted into a host bacterium lacking its own genetic material, the bacterium was able to successfully produce new cells.

Gene synthesis methods can even be used to construct new metabolic systems in living cells. For example, Jay D. Keasling constructed a new metabolic system in E. coli and S. cerevisiae in the mid2000s to produce artemisinin. An important component in some anti-malarial drugs, Keasling’s construct reduced the cost of artemisinin production by tenfold, showing that gene synthesis has a vast amount of potential in medicine and countless other fields.

Gene synthesis was first successfully conducted in 1972 on a yeast tRNA by Har Gobind Khrona and associates.Since 1972, the technology and mechanisms of gene synthesis have flourished in order to successfully obtain the genetic sequence specific to a wide range of research topics.Gene synthesis offers many major advantages over the traditional gene cloning methods: molecular cloning and polymerase chain reaction (PCR).These two traditional methods have two major restrictions,the first is that these methods are only capable of gene amplification and cloning, as opposed to gene synthesis.The other restriction molecular cloning and polymerase chain reaction have is that the preexisting genetic sequence of interest must be obtained and then amplified.The important advantage gene synthesis offers is that the genetic code you wish to study does not need to be physically obtained in order to be amplified.The technology currently allows us to synthesize “de novo” sequences.This means that a sequence of interest,whether it is viral DNA or a cancer cell’s mutation in a specific gene, can be synthesized without the physical copy of the gene being present.Without the physical copy of the gene of interest being needed,the necessary amount of time and money is drastically reduced when compared with traditional methods of cloning, amplification,targeted mutagenesis,and gene target knockouts.Due to this advantage gene synthesis has been becoming more and more popular in various fields of research.

Due to the ease of use and cost effective mechanism, gene synthesis has been used countless amounts of times in varying fields of scientific research. Gene synthesis has most notably and recently been used to conduct viral research in order to produce safer, more effective DNA vaccines. It has also been used to better understand mechanisms for cancer cell metabolic regulation. Gene synthesis is currently being used for targeted gene therapy, and other topics of interest that seemed almost impossible 30 years ago. In addition to the research that has already been conducted, gene synthesis offers a myriad of possibilities of application ranging from gene regulation to better understanding evolution and antibiotic resistance. Gene synthesis makes it possible to build variant libraries, genes, operons, increase the function of proteins, and even test all the orthologs of a particular gene.

The advantages gene synthesis offers are seemingly endless.The main advantage that gene synthesis has to offer is the ease of synthetically manipulating or cloning a gene of interest with or without the physical copy of the gene itself.Synbio Technologies offers, through our Syno^®DNA Platform, an extremely accurate, time efficient, and cost effective mechanism to analyze your sequence of interest and successfully synthesize the sequence.All sequences constructed by Synbio Technologies are verified using Sanger sequencing, and are guaranteed to be 100 percent identical to your sequence of interest.In addition to the high quality synthesized sequences, Synbio Technologies offers competitive prices on gene synthesis for a wide range of gene lengths up to 100 kb.With the competitive prices and accuracy, on top of the additional money and resources needed for PCR and molecular cloning, it is clear that gene synthesis is the more effective mechanism to utilize when conducting certain types of genetic research.

Gene synthesis has revolutionized genetic research over the past twenty years by allowing the convenience of DNA storage for a wide range of genome sizes. These libraries have allowed researchers to archive the genes of interested and be able to obtain each gene with relative ease at his or her leisure. Another great advantage ofgene synthetic libraries is the convenient location of the genetic foundation of the organism at your disposal. The genetic foundation can either be stored as the entire genome or more specifically the coding regions of the genome. Gene libraries are mainly broken down into two distinct categories: genomic libraries and cDNA libraries. The two have the same principles and similar output, but are created in different ways and have many different aspects.

First, the genomic libraries is representative of the organism’s entire genome, making the library quite large. The process of creating a genomic library is as follows: first the selected DNA is isolated from the cells by a specific restriction endonuclease. The resulting DNA fragments are then inserted into a selected vector using DNA ligase. Once the DNA is located within the vector, the vector is inserted into a bacteriophage to be amplified. After amplification, the cloned DNA is then isolated again and inserted into a genomic library. This process is then repeated until the entirety of the organism’s genome is isolated, amplified, isolated again, and inserted into the genomic library. Since it is the entire organism’s genome is located in these libraries, including both coding and noncoding regions, these libraries can become quite large. Genomic libraries offer many advantages, such as being able to study gene regulation, or off target effects of a particular mutation. The large amounts of data allow researchers to better understand how mutations, located outside of the coding region of a gene, affect the organism. This extra information is essential to better understand certain mutations, but the large amounts of data can sometimes be problematic. In conclusion, genomic libraries offer more information about the organism, but in turn are more laborious to create and are much larger in size when compared to cDNA libaries.

Second, cDNA library is representative of the organism’s exonic regions, meaning that only the coding regions of the genome are recorded and stored in the library. The process of creating a cDNA library is as follows: mRNA is isolated from a cell of interest and collected. Reverse transcriptase is then used to generate the double stranded cDNA corresponding to the mRNA sequence. Once this is complete, the DNA ends are cleaved to become single stranded and is inserted into a selected vector using DNA ligase. The vector is then inserted into a bacteriophage and amplified. After amplification, the cDNA is isolated and collected in the cDNA library. The cDNA library is much smaller than that of genomic libraries as it only represents the exonic portion of the organism’s genome. This small size is not always problematic, cDNA libraries offer a unique approach to study variant mutations present within coding regions of a gene. In addition to studying variant mutations located within a gene, cDNA libraries also offer a more convenient approach to study protein function, interaction and expression.

Whether you are interested in studying an organism with a very small genome or one as complex and large as a human genome, these two types of libraries are one of the best ways to store data. At Synbio Technologies we offer gene synthesis service verify the genes you are interested.A library, either constructed by you or our team of experts, will be generated for a competitive price and in an efficient timeframe. We offer virtually any library type to fit your need, ranging from scanning, to triplet codon, to modular. In addition to this Synbio Technologies also offers error-free gene sequence with a 100 percent guarantee of the generated sequence quality. This also allows us to create a synthetic gene library, generated by genes of interest without the physical copy of the gene being needed. Synbio Technologies offers many quality assessments to make sure that your gene library is exactly how you designed it to be and is as effective as possible. With the high quality library Synbio Technologies offers, and at a competitive price, your library will be synthesized and be ready to analyze with ease and convenience.