How is DNA and Gene Cloning Transforming Field of Regenerative Medicine?

When we hear the word “cloning,” the first thing that comes to our mind is recreating an entire organism, such as Dolly the Sheep. However, cloning something simply means developing a genetically exact copy of it. In molecular biology labs, gene cloning is commonly used by researchers to develop copies of a specific gene for downstream applications such as genotyping, sequencing, and mutagenesis.

DNA and gene cloning holds the key to understanding the blueprint of life itself. In this blog post, we’ll take you on an enlightening journey into the fascinating world of DNA, uncovering the steps and technologies behind the revolutionary technology. Also, we’ll delve deep into the uses and advancements in DNA and gene cloning. Read on!

What is DNA Cloning?

DNA cloning refers to the process of making several identical DNA copies of a specific DNA segment. In a typical DNA cloning procedure, enzymes are used to insert the specific DNA fragment into a circular DNA piece called the plasmid. The insertion, which is carried out using restriction enzymes and DNA ligase, results in the production of a molecule of recombinant DNA.

The next phase involves introducing the recombinant plasmid into bacteria. As the bacteria grow, they clone the plasmid and pass it on to their offspring, thereby producing multiple exact copies of the plasmid. These copies are referred to as clones.

Step of DNA and Gene Cloning

DNA cloning has a wide range of applications. For instance, researchers often use DNA and gene cloning services to synthesize a protein in bacteria. The basic steps involved in DNA and gene cloning are:

Cutting and Pasting DNA

How can DNA pieces from varied sources be glued together? A typical way of cutting and pasting DNA makes use of two types of enzymes. These are restriction enzymes and DNA ligases.

A restriction enzyme is a DNA-cutting enzyme that cuts the DNA at specific target sequences. Several restriction enzymes develop cuts with short overhangs. If in case two molecules tend to have identical overhangs, they can stick together. However, a DNA ligase is needed for the matching overhangs to produce an unbroken DNA molecule.

Bacterial Transformation and Selection

The transformation process involves the introduction of recombinant plasmid into bacteria such as E.coli. During transformation, the bacteria are subjected to very high temperatures that compel them to take foreign DNA. The plasmid carries an antibiotic-resistance gene that helps the bacteria survive in the presence of a specific antibiotic. Thus, the bacteria that took up the plasmid can be selected on nutrient-containing antibiotics. Here, the transformed bacteria survive, whereas the other ones die.

Protein Production

The final step involves growing a large culture of plasmid-containing bacteria. Here, the bacteria are given a chemical signal that prompts them to target the protein. After protein production, the bacteria are split open to release it. Finally, the protein is purified and isolated from other cell content.

Uses of DNA Cloning

DNA molecules built through cloning techniques find several applications in molecular biology. Here are some use cases of DNA cloning:

Biopharmaceuticals: DNA cloning is used for making human proteins with biomedical applications. One example is the development of insulin using bacteria called E. coli. Another example of recombinant protein is tissue plasminogen activator (tPA), which facilitates the breakdown of blood clots. Proteins like tPA are typically produced in bacteria.

Gene Therapy: In some genetic disorders, patients typically lack the functional form of a particular gene. Gene therapy is a technique that aims at producing therapeutic effects through the manipulation of gene expression or by changing the biological properties of living cells. For example, plasmids built using DNA cloning can reduce the deterioration rate of lung function in cystic fibrosis patients.

Gene Analysis: DNA and gene cloning is used by biologists in basic research labs to develop artificial versions of genes that help them understand the functioning of normal genes in an organism.

Types of DNA and Gene Cloning Services

Here’s an overview of the most common types of DNA and gene cloning services:

Gene Synthesis: Gene synthesis, also known as artificial gene synthesis, refers to a group of methods used in synthetic biology to construct and assemble genes from nucleotides de novo. Unlike DNA synthesis, gene synthesis doesn’t need template DNA. Thus, any DNA sequence can be synthesized in a lab.

Custom Cloning: Custom cloning is the process that develops recombinant DNA molecules and transforms them into a host organism. The custom cloning process typically includes a DNA fragment of interest and a plasmid backbone.

Subcloning: Subcloning is a technique in molecular biology that involves moving DNA inserts from one vector to another for study. The insert sequences may be taken from any organism, including the mRNA, genome, or previously cloned DNA segments.

Latest Developments in DNA and Gene Cloning Services

The market for DNA and gene cloning services is highly competitive. Top industry participants such as Thermo Fisher Scientific, GenScript, Agilent Technologies, Takara Bio, Lonza Group, and Genwiz focus on continuous innovation to introduce advanced services in the market. Below are the latest developments in the market:

• In June 2024, GeneScript Biotech Corporation launched the GenScript FLASH Gene service. As an ultra-fast sequence-to-plasmid (S2P) service, GenScript FLASH Gene service has reduced the delivery time from sequence to plasmid construct to just four business days.
• In November 2023, Twist Bioscience launched its Twist Express Genes Service. With the new service, Twist can deliver the NGS-verified clonally perfect genes to its customers in about half the time.

Improving Genetic Research Capabilities

To conclude, DNA and gene cloning represent profound advancements in genetic science, providing remarkable potential across various sectors. By allowing scientists and researchers to manipulate specific genes, DNA cloning paves the way for developing groundbreaking treatments for genetic disorders and getting a deeper understanding of sophisticated biological processes. The future ahead promises exciting developments in the realm of DNA and gene cloning services.

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