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Gene Synthesis: An Overview Of Its Use In Biotechnology

Oct 7th 2022

Gene Synthesis: An Overview Of Its Use In Biotechnology

Gene synthesis is integral to molecular biology, generating probes for live science industries. Such industries include drug discovery, vaccines, gene therapy, tissue replacements, human genomes, disease detection research, and improved nutritional quality in food.

Classic genetic methods used a few decades ago only facilitated the modification of one molecule at a time. However, gene synthesis companies' next-generation sequencing services facilitate the production of recombinant genes, mutated genes, and novel DNA via custom gene synthesis.

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Gene synthesis is the baseline for gene synthesis services like peptide synthesis, protein production, antibody production and sequencing, and ELISA kits production. Learn more below.

What Is Gene Synthesis?

A gene is the primary genetic unit carrying genetic information or hereditary traits transferable from parent to offspring. It features nucleotides that bind together to form a DNA molecule. Genes occur naturally as nucleic acid sequences in the chromosomes present in the human cell. However, gene synthesis creates synthetic gene material nucleic acid sequences as base pairs.

Gene synthesis services allow researchers to optimize or modify genetic components to boost the target gene’s traits or characteristics for use as probes in various live sciences research projects.

Traditional gene synthesis techniques included mutagenesis or introducing a mutagen (mutation-causing agent) in a genetic code. However, mutagenesis only altered single base pairs in an entire gene sequence and, as such, was not viable for projects like custom DNA synthesis or custom gene synthesis.

The second classic gene synthesis option was traditional cloning with restriction enzymes. A restriction enzyme is a bacteria-generated protein that cleaves or cuts DNA sequences along target sections to create DNA fragments like genes.

Although traditional cloning has multiple advantages, including low labor intensity and diverse expression systems, like the E coli protein expression system that DNA synthesis companies use in rendering protein expression and purification services.

However, the classical custom gene synthesis technique had disadvantages, including possible sequence constraints resulting from restriction sites. Other disadvantages included undesirable recombination sequences in protein production services and a vector structure that may affect downstream services like recombinant protein production and antibody sequencing services.

The first historical mention of gene synthesis was in 1972, when Gobind Khorana, an Indian chemist, developed the first in vitro functional synthetic gene. He used multiple enzymes to link DNA fragments into a gene.

Fast-forward to 2022, and advantages in biotechnology make it possible to generate an entire synthetic genome from gene synthesis, create non-natural difficult-to-extract natural gene sequences, and novel sequences that do not occur naturally.

Most gene synthesis companies offer custom DNA synthesis services and can generate 150-200 kilobase pairs (kb) synthetic gene lengths using techniques like the solid-phase peptide synthesis protocol. Moreover, some gene synthesis companies use ligation techniques to generate longer DNA strands with thousands of base pairs.

Therefore, besides transferring genetic traits to offspring, the genes in DNA molecules are vital components in peptide synthesis and protein production services like antibody production. Consequently, gene synthesis is a standard product for gene synthesis companies.

However, such companies build on gene synthesis services by offering line services. Such services include peptide synthesis, custom protein synthesis, custom antibody service, custom antibody expression, Protein expression services, antibody sequencing services, and ELISA kits production.

Custom gene synthesis companies have varying costs depending on the length and complexity of the peptide, expression systems, and the customization level for custom gene synthesis. Other DNA synthesis companies charge clients per base pair.

Nonetheless, gene synthesis revolutionized biotech processes, including protein production services, protein expression service, solid-phase peptide synthesis protocol, antibody production, and peptide synthesis services. Below is an overview of gene synthesis benefits to DNA synthesis companies and recombinant protein production services in the life sciences.

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Gene Synthesis Advantages

In vitro gene synthesis services have multiple advantages over traditional gene synthesis techniques. Below are the top reasons why DNA synthesis companies offer gene synthesis services as a primary product.

1. Quick Turnaround Time

Classic gene synthesis methods have a slow turnaround time, taking service providers months to deliver the target gene required for research and industrial purposes. As a result, research projects would stall, and others would shut down altogether due to the timelines it took DNA synthesis companies to generate genes and DNA products.

In contrast, contemporary gene synthesis methods, like the solid-phase peptide synthesis protocol, have a significantly lower turnaround time for the gene synthesis protocol. Although the turnaround time varies depending on the gene length and subcloning difficulty, the industry average turnaround time for gene fragments is 6-9 days, while the turnaround time for clonal genes is 14-21 days.

Moreover, some gene synthesis services can offer complimentary services, including free standard vectors for subcloning in protein expression systems. Also, the final gene product delivery as a lyophilized plasmid takes less than 24 hours within US borders. Therefore, researchers can order gene products online from DNA synthesis companies.

2. High Accuracy Levels

Most gene and custom DNA synthesis service providers guarantee 100% gene sequence accuracy. Custom DNA synthesis companies use PCR (polymerase chain reaction) methods to ensure gene product accuracy.

PCR is the standard method for generating exact copies of a target DNA or gene molecules into millions of molecules. High accuracy in gene replication is crucial to facilitate consistent research results or product delivery in applications like antibody production, protein expression, recombinant protein production, ELISA kits production, and custom protein production.

Although multiple PCR methods are available, dial-out PCR is the standard industry technique for ensuring gene product accuracy. Moreover, some gene delivery services provide complete gene sequencing quality control data. Such data includes a quality analysis report, construct map, the entire sequence, chromatograms, and an alignment file.

3. Affordability

The ubiquity in gene synthesis services makes gene product services like peptide synthesis, custom protein synthesis, custom antibody, and protein expression and purification services more affordable. Moreover, most service providers have a “no gene,’ no charge” policy, protecting researchers and commercial biotech stakeholders from potential losses from low-quality genes and other DNA products.

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Gene Synthesis Protocol/ Standard Workflow

Gene synthesis is a step-by-step process that begins with an oligonucleotide (short DNA or RNA fragment) and ends with a gene plasmid ready for delivery and use in an industrial or lab setting.

The two primary methods used in gene synthesis are polymerase cycling assembly (PCA) and gene synthesis by ligation. Although each method has advantages and disadvantages, both techniques follow a similar protocol to maximize the target gene yield. Below is an overview of the gene synthesis protocol leading to a high-quality, active gene molecule.

1. Chemical synthesis of oligonucleotides

Gene synthesis begins with the chemical synthesis of an oligonucleotide. The step is crucial for recombinant protein production processes like custom antibody service, custom protein synthesis, custom protein expression, peptide synthesis service, and custom protein production.

Chemical oligonucleotide synthesis occurs in the 3’ to 5’ direction and entails using the solid-phase peptide synthesis protocol to attach a chemically-modified nucleoside to a polymeric support. The polymeric support holds a growing oligonucleotide chain, which undergoes deprotection, as is the case with recombinant protein production and the peptide synthesis service.

2. Oligonucleotide Annealing By Ligation or Polymerase Reactions

The newly-formed oligonucleotide undergoes annealing as the second step in the gene synthesis protocol. Annealing entails heating a molecule and then cooling it slowly to facilitate hybridization. Hybridization is the fusion of two atomic orbitals to form a new orbital.

As earlier stated, an oligonucleotide is a DNA fragment and features the double-strand structure typical of DNA molecules. Therefore, the heating process in annealing splits the oligonucleotide into two strands.

On the other hand, the cooling process merges multiple single-strand oligonucleotides into a single long chain. However, care is necessary to prevent E-coli protein aggression or property alteration during annealing.

PCA and synthesis by ligation are the primary processes used in oligonucleotide annealing. Custom protein production processes like custom antibody production and custom protein expression dictate the ideal annealing method.

3. Gene Sequence Cloning Into Vectors

Cloning is a process entailing replicating pieces of DNA into multiple copies. Oligonucleotide annealing facilitates the formation of a complete gene sequence. Therefore, the next step is cloning that gene for research and industrial purposes like peptide synthesis service, custom antibody service, protein expression service, and recombinant protein production.

A cloning or plasmid vector is a piece of DNA replicating independently from the cell and is the primary replication machinery in gene synthesis. Although natural cloning vectors like E-coli protein expression systems are available, synthetic plasmids have fewer steps of protein purification. Therefore, such plasmids are better options for protein expression and purification services.

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4. Clone Screening

Clone screening is necessary to identify and isolate acceptable target genes from the E-coli protein expression system. ELISA kits are ideal clone screening tools, helping to detect the target gene in a biological sample.

Gene product service providers typically partner with ELISA kit manufacturers to acquire ELISA tests for effective clone screening. However, other screening methods like chromatography are available besides the test kits by ELISA kit manufacturers.

5. Sequencing Analysis

Gene sequencing entails determining the genetic composition of a gene or DNA by examining the base pairs. The process utilizes sequencing primers, including gene-specific primers, to reveal the gene sequence.

Additionally, gene sequencing goes hand-in-hand with gene analysis or study. Gene product research studies on antibody production service, antibody specificity, monoclonal antibody sequencing, and how to make polyclonal antibodies can run their projects using sequenced genes retrieved from E-coli protein expression systems.

6. Error Correction

Replication errors like base substitution, deletion, and incorrect insertions are common in gene sequencing. Although such errors negatively impact analysis processes like antibody specificity and monoclonal antibody sequencing, error correction is possible to ensure correct plasmid placement.

7. Plasmid Purification and Lyophilization

Different research projects, like antibody production service and antibody sequencing services, require different gene purity levels. However, synthetic genes require minimum purification, meaning the protein purification service is not mandatory. Moreover, some service providers offer the protein purification service as a complimentary service.

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Gene Synthesis Applications In Biotechnology

As earlier stated, gene synthesis is a foundational process for most biotech applications. Below are the top gene synthesis applications for research and industry.

1. Antibody Production

Antibody production entails the production of antigens immunized into a host to generate target antibodies. Although antibody production is natural, gene synthesis has multiple advantages, including preventing side reactions and toxicity to the host. Second, synthetic genes have enhanced properties, including a higher antibody specificity and low E-coli protein aggregation likelihood.

Custom antibody service for monoclonal antibody sequencing services and steps to make polyclonal antibodies begin with gene synthesis. Moreover, gene sequencing lowers the custom antibody cost for monoclonal antibody sequencing. Also, research into making polyclonal antibodies has contributed to developing potent polyclonal antibodies with two binding sites for enhanced antibody specificity.

2. Recombinant Protein Production

Recombinant protein production is a multifaceted process featuring elements like protein expression service, antibody production service, custom protein production, antibody sequencing services, custom protein expression, antibody production service, and custom antibody production.

Gene synthesis is the genesis for the above processes, facilitating solid-phase techniques and the steps for protein purification. Additionally, recombinant protein production utilizes high throughput techniques to generate high-quality proteins for custom antibody production in drug discovery, therapeutics, and genetic modification for food security. According to one report, 239 FDA-approved therapeutic proteins and peptides are available for clinical use in the US.

3. ELISA Assay Test Kit Production

Enzyme-linked Immunosorbent Assay (ELISA) kit is an assay kit featuring peptides embedded on multi-well plates. So, how does ELISA work? ELISA kit manufacturers design the kits with antibodies to measure the presence and volume of targeted proteins and antibodies.

The kits have multiple applications in disease detection and diagnosis. Also, ELISA kit manufacturers optimize the kits for protein specificity in the protein purification service. Most gene product service providers bear the ELISA test kit cost, lowering custom antibody costs.

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Gene synthesis revolutionized biotechnology by facilitating research to develop optimized genes to improve the quality of life within and outside medical fields. Synthetic genes enable high throughput gene product generation for the agricultural, cleaning, pharmaceutical, beauty waste treatment, and food processing industries. Therefore, gene synthesis is biotechnology's future, and researchers are constantly working on new methods to make the process more cost-effective and efficient.

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