Messenger RNA (mRNA), is a single-stranded molecule in cells that carries codes from the DNA in the nucleus to the sites of protein synthesis in the cytoplasm (the ribosomes). With the major technological innovation and research investment, mRNA has become a promising therapeutic tool in in a variety of applications, including cancer immunotherapy, infectious disease vaccines, protein substitution and cellular genetic engineering. mRNA-based therapeutics hold the potential to cause a major revolution in the pharmaceutical industry because they can be used for precise and individualized therapy, and enable patients to produce therapeutic proteins in their own bodies without struggling with the comprehensive manufacturing issues associated with recombinant proteins. Compared with the current therapeutics, the production of mRNA is much cost-effective, faster and more flexible because it can be easily produced by in vitro transcription, and the process is independent of mRNA sequence. In addition, with the ease of rapid, large scale Good Manufacturing Practice-grade (GMP-grade) mRNA production, mRNA is ideally poised potential drug candidate that will meet the needs of gene therapy, cancer therapy as well as vaccination and so on.

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The Rosa26 locus on mouse chromosome 6 is often used to integrate transgenic structures to achieve ubiquitous or conditional gene expression in mice and is considered to be one of the ideal locations for knockin targeted genes. The DNA fragments placed at the Rosa26 site are very safe and stable. Using the classical gene targeting method, obtain a gene knockin mouse at Rosa26 always takes one year or more. With the application of CRISPR/Cas9-mediated knockin technology, large DNA fragments can be accurately and efficiently inserted into the Rosa26.

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Genomic engineering in cell lines is a versatile tool for studying gene function, designing diseases models, biopharmaceutical research, drug discovery and many other applications. CRISPR (Clustered Regulatory Interspaced Short Palindromic Repeats)/Cas9 systems is a newly developed yet the most popular method for genome editing. It has been widely used in current biology, functional genome screening, cell-based human hereditary disease modeling, epigenomic studies and visualization of cellular processes.

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