Gene synthesis refers to the base-by-base chemical synthesis of DNA strands. This technology is becoming an essential tool in recombinant DNA technology and replacing the traditional cloning and mutagenesis procedures.
Yet, there can be many unanswered questions in your mind. With this article, we aim to answer some questions that you must know about gene synthesis!
1. What is the Process of De-Novo Gene Synthesis?
In gene synthesis, single-stranded oligonucleotides are designed, synthesized, and assembled using different procedures. Subsequently, the assembled full-length gene is cloned into a vector. Finally, various techniques like restriction digestion and Sanger DNA sequencing are used for sequence verification.
The basic steps of gene synthesis are:
- Selection of gene of interest
- Sequence optimization
- Oligo design and synthesis
- Gene assembly
- Cloning in the vector of choice
- Sequence verification and error correction
- Preparing synthetic DNA for downstream applications
You can partner with gene synthesis companies for quick turn-around time, cost-effective, and highly efficient solutions.
2. What Can Be Synthesized Using Gene Synthesis?
With gene synthesis, you can generate recombinant or completely novel DNA sequences without a template. Apart from DNA sequences, you can also synthesize oligonucleotides containing modified bases. In addition, RNA or chimeric DNA-RNA backbones can also be synthesized. For instance, you can synthesize cDNA for heterologous or overexpression.
3. How does Gene Synthesis Differ from Molecular Cloning?
It’s crucial to understand that gene synthesis and molecular cloning are not the same. Unlike molecular cloning, gene synthesis does not require a template. Instead, researchers can specify a desired sequence and custom build it.
Gene synthesis is faster and less costly than molecular cloning. Yet, you can combine both technologies to make the synthetic products ready to use in research applications.
4. What is the Significance of Codon Optimization?
Codon optimization implies modifying the DNA sequence without altering the amino acid sequence to enhance efficient heterologous protein expression. Numerous studies reveal that codon-optimized genes generate more and high-quality proteins than original genes.
There are multiple ways to modify the gene.
- Add or withdraw restriction sites to facilitate gene manipulations.
- Add ribosome binding sites, linkers, and introns.
- Modify N- or C-terminal tags for conjugation and affinity purification.
5. Why is Gene Synthesis a Groundbreaking Tool for Research?
Gene synthesis is accelerating research across various disciplines by providing critical advantages over more laborious traditional molecular cloning methods.
- Genetic Engineering
With gene synthesis, researchers can generate novel biological systems and functions with gene-editing technologies like CRISPR.
- Antibody Discovery
Codon-optimized gene libraries and constructs allow you to expedite candidate screening for targeted therapeutic antibody engineering.
- Gene Therapy
Gene synthesis allows you to clone synthetic therapeutic genes into custom viral vectors efficiently. In addition, it will optimize the specificity of gene delivery and expression.
- Structural Biology
You can use recombinant protein expression for structure determination in gene-to-protein channels.
- Protein Engineering
With codon-optimized synthetic genes, you can boost protein expression levels for rational design and directed evolution.
Over the last few decades, our capability of synthesizing genes and assembling them has skyrocketed from less than 100bp to more than 1,000,000bp. As a result, Gene synthesis protocols and applications profoundly impact vaccine design, proteomics, and metabolic engineering.
Moreover, the continuous improvements and developments in the field make gene synthesis a powerful and error-free tool in molecular biology.