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Gene synthesis is a term used in synthetic biology to describe a set of methods. These methods are for making and putting together genes from scratch. Unlike DNA synthesis in living cells, gene synthesis doesn’t need a template, meaning that we can make almost any DNA sequence in the lab.  

However, being such an advanced process, there are various unknown facts about gene synthesis. The following part offers vital information regarding gene synthesis and related facts. 

 What is Gene Synthesis? 

Gene synthesis refers to the base-by-base chemical synthesis of a DNA strand. It does not need a template strand, unlike the DNA replication in cells. However, it involves the addition of nucleotides to a single-stranded molecule. This process serves as a template for the construction of a complementary strand. 

The area of synthetic biology started with the core technology of gene synthesis. We can synthesize any DNA sequence, including sequences that don’t exist in nature. Also, we can clone synthetic DNA into expression vectors and use them where natural DNA is necessary. 

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 Gene synthesis technology has changed the method of research in biology. Scientists don’t have to change one gene at a time anymore. They can now make and change entire genomes and cells. To speed vaccine research, we can quickly create new viral genomes.  

To better understand the basic needs of life, we can make synthetic genomes and make artificial cells. As a result, life science experts from various fields can do more intense research. 

 Four Unknown Facts About Gene Synthesis 

There are several pieces of information about gene synthesis on the internet today. However, not every piece of information is reliable and valid. Moreover, many are unknown to what gene synthesis holds. Here are four unknown facts about this complex process. 

  1. Codon Optimization is Not the Only Benefit of Gene Synthesis 

 Codon optimization describes experimental methods for improving the codon composition of a recombinant gene without changing the amino acid sequence.  

Most scientists use gene synthesis to increase their gene expression in heterologous systems. Codon usage adaptation can help with this.    

 However, there are many more advantages to employing gene synthesis. You can order your DNA or RNA sequence with gene synthesis; all you need is the silico sequence.  

Another benefit of gene synthesis is having quick and dependable access to cDNA sequences. This was only possible through time, labor, and expensive cDNA synthesis.  

Also, no RNA extraction or RACE is necessary to get flawless full-length cDNA while using gene synthesis. You can also subclone your own expression vector. 

  1. We can Synthesize Complete Plasmids
  2. It is possible to synthesize genes of any length, including complete plasmids. During the gene synthesis process, overlapping DNA oligonucleotides are joined together to form lengthy pieces. These long pieces then combine to create a sizeable synthetic gene. It is even possible to generate whole plasmids by adding a circularisation technique. 

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 Here are some benefits of synthesizing a plasmid: 

  • The vector’s nucleotide sequence matches the host organism to control the expression levels of genes in antibiotic resistance. 
  • You can make your own multiple cloning site for your project. 
  • You can choose the selection markers, replication origin, restriction sites, and other features. 
  • If you want to use your plasmid for business, there are no licensing issues to deal with.
  • We Can’t Optimize a Non-Coding Sequence

 Due to DNA’s repetitive nature, more than one triplet codon can code one amino acid. For example, CGT, CGC, CGA, CGG, AGA, AGG, etc., can code arginine. The number of these codons varies based on the species.  

So, we can optimize the DNA sequence to have the same number of codons as the animal in which it will be expressed. When the gene’s high GC and repeat areas are optimized, they are also less. 

 However, if the gene to be optimized codes for a protein, this type of optimization is possible, as the codons that make up the protein can be changed.  

So, optimization is not possible because non-coding DNA stretches don’t have codons that code amino acids. 

  1. We can Subclone Inserts Into a Vector
  2. Even if your insert lacks restriction enzyme sites from your vector’s multiple cloning site (MSC), you can still subclone it into the vector. 

You need one or two restriction enzymes that target the MCS to cut your vector. Your gene insert can then be PCR amplified with primers. This is to generate vector-homologous sections at the ends of the PCR result.  

These homologous sections are subsequently subcloned into your vector using SLIC, which means sequence and ligation independent cloning.  

You don’t need to include any restriction sites in your insert, as the PCR product isn’t affected when you put it into a new cell. SLIC is as quick and efficient as typical restriction enzyme site subcloning.  


Gene synthesis allows for the rapid generation and manipulation of genetic sequences, opening up vast possibilities for new biological functioning. However, people have accepted several myths about gene synthesis, and the false information keeps spreading out to more. This is why we have gathered some facts that people have misunderstood in the past. Moreover, gene synthesis is essential, so we need to have the correct information about it at hand. 


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