This post introduces the latest technology of engineering phages and how they are applied to develop phage-based therapy.
Updated Phage Engineering Techniques Witness Bacteriophage Therapy Market Growth
Phages, as the most abundant organisms on the earth, once were found to be a powerful weapon to selectively target and kill bacteria, though their reputation was overtaken by antibiotics in a period. Now, the increasing spread of antibiotic resistance brings phage therapy back to the historical stage of microbiology, attracting researchers to unlock the potential of phage therapy from all aspects.
Reports from different market research agencies indicate that the global bacteriophages therapy market is expected to gain market growth at a CAGR of 3.7% in the forecast period of 2021 to 2027, with an impact of COVID-19 and recognition of phage genetic engineering. Apart from natural phages that are usually discovered as bacterial sensing agents in pathogen detection, modified phages in phage therapy now also take an important part in the bacteriophage therapy market. Currently, a growing number of life science companies and researchers are working on phage-based new therapeutic projects.
Creative Biolabs, a leading life science company dedicated to comprehensive phage-based services, has accumulated extensive experience in the development of different engineered phages using currently updated techniques of phage engineering.
Homologous Recombination Technique
One of the most frequently used bacteriophages engineering methods is homologous recombination in bacterial hosts, which involves the physical exchange of strands between homologous or nearly homologous DNA molecules. By inserting, replacing, or removing genes from phage DNA sequences, the homologous recombination technique allows modification of phages for different research interests.
Bacteriophage Recombineering Electroporated DNA Technique
Another updated approach to phage engineering is bacteriophage recombineering of electroporated DNA (BRED), which can be used to delete, insert, and replace genes as well as to generate point mutations in phage genomes. BRED aims to obtain engineered phages by increasing the recombination frequency (up to 10 to 15%) between a template and a bacteriophage.
In Vivo Recombineering
Different from other techniques, the in vivo recombineering method uses phage lambda as a tool for the engineering of other, less well-studied E. coli phages. With the control of the pL operon that takes part in general and site-specific recombination in E. coli cells, defective prophages carried by E. coli cells can be infected and engineered by the phage that is used as an in vivo recombineering tool. This technique allows about 0.5 to 2% yield of recombinant phages.
CRISPR-Cas-Mediated Genome Engineering
Due to unassigned functions existing in the majority of phage genes, CRISPR-Cas systems have also been harnessed for phage genome engineering. The action mode of CRISPR-Cas systems covers three main processes of CRISPR adaptation, RNA biogenesis, and CRISPR-Cas interference. Recent studies assured that CRISPR-Cas systems, including types I, II, and III, are powerful tools to facilitate phage genome engineering, with successful editions of three families of tailed phages such as Myoviridae, Siphoviridae, and Podoviridae.
Yeast-based Assembly of Phage Genomes
Large phage genomes bring difficulties for in vitro manipulation and in vivo manipulation also be hindered by the short existing time in bacteria. What's more, altering individual phage genes in bacterial cloning vectors using classical biology is not satisfying enough for scientists due to the slow and tedious process and possible toxicity to bacteria. All issues can be addressed by using yeast as an intermediate host for genetic manipulation. This yeast-based assembly technique has been successfully applied to capture and genetically modify the genomes of the coliphages T3 (38,208 bp), T7 (39,937 bp), and the Klebsiella phage K11 (41,181 bp).
Cell-free Expression and Synthesis of Bacteriophages
The efficacy of cell-free gene expression (CFE) systems has been improved to express DNAs composed of tens of genes encoding for complex self-assembly processes. A recent study demonstrated that infectious viruses and phages can also be synthesized in CFE systems.
About Creative Biolabs
Creative Biolabs is an industry leader in the field of phage research, equipped with advanced platforms for phage research and scientists who are qualified in experience, professionalism, and a vast reserve of knowledge. It's dedicated to providing top-tier services for phage research, including phage identification, small and large-scale phage production, phage purification, phage vaccine development, and phage therapeutics discovery.
Please find more possibility of phages at: https://phagenbio.creative-biolabs.com/
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