Cloning is a process used to produce genetically identical copies of biological entity. These copies are referred as clones. Since last decade, researchers are cloning wide range of biological material namely genes, cells, tissues, or entire organisms. Previously, researchers found that some plants and bacteria can produce genetically identical copies by a sexual reproduction. However, the species specificity reduces the chances of uptake of DNA from other species. Further, they tried to develop the identical clones using in vitro methods or laboratory conditions. It led to develop a new step in cloning experiment called transformation. This process includes direct uptake of DNA released into an environment by host cells and simultaneously integrates that DNA with host genome for recombination.
Naturally, many bacteria have an ability to take up DNA from environment. The DNA provides nutrients to these natural competent cells, while recombination leads to new genetic evolution. This ability to take up external DNA naturally is referred as natural competence. “Meticulous Research” in its latest publication states that, the “global Competent Cells Market for is expected to grow at a CAGR of 10.0% from 2018 to 2023 to reach $2,318.6 million by 2023.”
This growth is primarily driven by growing demand for recombinant proteins & molecular cloned products, advancement in molecular cloning research with emergence of new technologies, and rising government initiatives and funding for life science research.
Advancements in molecular cloning have significant impact on competent cell market. Molecular cloning refers to the isolation of a DNA sequence from any species, insertion of that gene into vector for propagation without altering the genetic make-up. It includes various steps such as isolation of target DNA fragment; ligation of insert into cloning vector, creating recombinant molecule; transformation of recombinant plasmids into suitable host for propagation; and screening & selection of hosts containing target DNA.
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Traditionally, researchers were using all basic principles for creating and propagating recombinant DNA in bacteria. However, these all were found to be inefficient. For instance, restriction enzyme preparations were unreliable due to non-standardized laboratory protocols; in addition, plasmids for cloning found to be cumbersome, difficult to work, and limited in DNA insert. This led to incline researchers towards improvements in techniques and tools used for molecular cloning. Researchers recognized the need for general cloning plasmid with unique restriction sites for cloning in foreign DNA and the expression of antibiotic resistance genes for selection of transformed bacteria. This led to develop the general cloning plasmids. Although, antibiotic selection in plasmids restricted the growth of non-transformed host, these plasmids found could re-ligate without DNA insert and give antibiotic resistance. Thus, finding correct clone became difficult and time consuming. To overcome these drawbacks, researchers developed new techniques for selection of molecular clones, namely blue-white screening, where placement of multiple cloning sites (MCS) within Lac Z gene allowed to screen host cells with target DNA. In addition, previous work related to cloning was suffered due to lack of purity of enzyme preparation and lack of understanding of buffer requirements for each enzyme. Thus, pharma and biotech companies produced and commercialize restriction enzymes produced from a recombinant source. This enabled higher yield, improved purity, consistency, and lower pricing.
Researchers also developed different techniques for preparing vectors prior to ligation. This can be done by discovery of alkaline phosphatases, enzyme that remove 3´ and 5´ phosphate groups from the ends of DNA; thereby prevent self-ligation and increase recovery of plasmids with insert. In addition, the researchers developed polymerase chain reaction (PCR) to amplify target DNA by using primers to amplify both complementary strands of DNA. This technique reduced the issue related to amount of DNA in molecular cloning. This PCR was further evolved to Real Time PCR, multiplex PCR, and others so as to clone messenger RNA. After introduction of PCR, several different methods used for cloning PCR products. For instance, researchers introduce restriction sites onto the ends of the PCR product to allow directional cloning (insert into the vector after restriction digestion), blunt-end cloning (insert PCR products generated by polymerases that produced blunt ends). After this, overlap extension PCR was developed to assemble PCR products into one contiguous DNA sequence.
To improve the efficiency of molecular cloning, techniques such as TA cloning, LIC (Ligation independent cloning) cloning, and USER cloning were developed. TA cloning used property of Taq DNA Polymerase, the first heat-stable polymerase used for PCR. This helped to ligate PCR products into a vector that has cut and engineered to contain single T residues on each strand.
Further, LIC cloning allowed joining of DNA molecules in the absence of DNA ligase. This method further developed USER cloning, which allowed PCR independent of ligase and restriction enzyme cloning method. According to the NCBI, in 2016, researchers claimed that novel archetype may establish in synthetic biology techniques that can facilitate quicker assembly and iteration of DNA clones; thereby accelerating the progress of gene therapy vectors, recombinant protein production processes, and vaccine production by in vitro chemical synthesis of any in silico-specified DNA construct. Thus, advancement of these techniques improved molecular cloning from the cloning of a single DNA fragment to the assembly of multiple DNA components into a single stretch of DNA; thereby driving the competent cells market.
The report provides meticulous analysis of competent cells market on the basis of type (cloned competent cells, agrobacterium tumefaciens competent cells, and expression competent cells), treatment (chemically competent cells and electrocompetent cells), application (cloning, protein expression, others), end user (pharmaceutical & biotechnology industry, academic & research institutes, and contract research organizations), and geography.
The key players analyzed in the competent cells market are Thermo Fisher Scientific, Inc., Agilent Technologies, Inc., Bio-Rad Laboratories, Inc., Merck & Company, Inc., Illumina, Inc., Promega Corporation, New England Biolabs, OriGene Technologies, Inc., Zymo Research Corporation, Scarab Genomics, LLC, Meridian Bioscience, Inc., Delphi Genetics S.A., Qiagen N.V., Cell Applications, Inc., GenScript Biotech Corporation, and Takara Holdings Inc.
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