One of the fundamental ways to study the function of a gene is to void, knockout, its function and observe the resulting phenotype. This void of function, commonly known as mutagenesis, has been practice within the field of genetics for many years and has been proven effective countless times. A more commonly utilized method of mutagenesis, site-directed mutagenesis, is one of the most traditional methods of generating a knockout gene. The process of gene synthesis mutagenesis begins with the selection of a short primer. The primer must meet two important characteristics: it must be complementary to the sequence of interest, and it must contain a debilitating mutation that will leave the gene of interest incapacitated. The complementary nature of the primer is necessary for the primer to hybridize with the sequence of interest. The debilitating mutation is necessary in order to generate a knockout in the gene of interest. The single stranded primer is then replicated by DNA polymerase, allowing the amplification of the gene of interest which now includes the mutation specific to the primer. The mutated sequence is then inserted into a bacteria host cell via a viral vector and cloned. After amplification, the resulting sequence is then verified for accuracy. This process is laborious and may also not yield a high quality of resulting sequences. This inefficiency is due to the reliance of multiple different biological methods that are not easy to control. Over the past thirty years there have been multiple additional methods to generate mutated sequences but few with high efficiency. A new method, gene synthesis mutagenesis, has the recently become more popular as a result of its effective results and efficiency. Gene synthesis mutagenesis offers a unique approach to generating a genetic knockout within an organism. Instead of the normal site-directed mutagenesis, and somewhat hoping that the induced mutations will render the gene inactive, gene synthesis mutagenesis has the ability to generate any type of mutation within a gene. This gene synthesis is carried out by our Syno® Platform, which guarantees one hundred percent accuracy of the generated sequence. This guaranteed sequence will now contain a debilitating mutation, resulting in the creation of a knockout with little effect and high probability of success.
Gene synthesis itself has revolutionized the field of genetics. This unique technology offers the ability to synthetically engineer a gene of interest in order to be studied. Since this is possible, it is also possible to generate a gene of interest and insert mutations. This is the basic background of gene synthesis mutagenesis. Gene synthesis is the process of requested genetic sequenced in text format to the engineering of a physical copy of the requested sequence. This process has multiple steps of verification of the sequence of interest. These verifications and overall process can be altered in order to generate a sequence of interest that renders the previously functional gene inactive. Multiple different types mutations can be synthetically engineered, ranging from a frameshift mutation to a large deletion of an entire codon, both of which result in a gene damaged beyond repair. This gene can then be amplified, be either use of polymerase chain reaction or traditional subcloning, and then studied for the effects the knocked out gene has on the organism. The process of gene synthesis mutagenesis completely bypasses the reliance of DNA polymerase replicating the primer necessary in site directed mutagenesis. It relies solely on the ability to generate a mutation in text format, which can easily be done.
The connection between Synbio Technologies and gene synthesis mutagenesis is a simple one, we pride ourselves as being one the leading companies in the gene synthesis industry. On average we synthesize over 10 million base pairs a month. This process is carried out by individuals who on average have been working in this field for 10 years or longer. With this experience and constant business we have been able to perfect the pipeline from original text format to engineered product through our Syno Platform. This format is then slightly altered to accomplish gene synthesis mutagenesis requested by the customer. Instead of using the Syno Platform to synthetically engineer a functioning gene of interest, we engineer a gene of interest with a debilitating mutation. Since Synbio Technologies is one the premier companies in the gene synthesis industry, we can also be considered to be one of the leading companies in the gene synthesis mutagenesis industry.
Gene Synthesis Related Services
Synbio Tenchologies can also design sequencing with codon optimization software -NGTMCodon Optimization Technology at no cost.