This year Thermo Fisher Scientific has made impressively swift adaptations to the rapidly changing landscape of genetic engineering. By doing so, they have positioned themselves to cater to scientists’ needs at the forefront of research by introducing new products and services for genome modification as well as on-demand gene and protein synthesis.
The most notable addition to their product-lineup is their recently perfected suite of tools for CRISPR-based genetic engineering. They offer services for every level of research and at every step of the process from design to delivery and detection.
Thermo Fisher Scientific developed the online GeneArt CRISPR Search and Design Tool to allow users to take advantage of their pre-designed gRNAs and advanced algorithms for de novo gRNA designs. Not only do they have 600,000 predesigned CRISPR gRNAs for targeting human and mouse genes, for any particular gene you choose, the design tool suggests the top 6 sequence designs and gives a list of potential off-target effects for each. On the off chance you want to target a gene not included in their library, their Design Tool will give you up to 25 gRNA suggestions once again with each sequence’s potential off-target effects.
Thermo Fisher’s CRISPR delivery system comes in four forms: all-in-one plasmid vectors, CRISPR mRNA, purified Cas9 protein, and Lentiviral libraries each paired with its optimal cell culture reagents and delivery method. The reason for such a diverse set of delivery options is due to every cell-line having a unique transformation and cleavage efficiency for each method. Thermo Fisher has compiled a large list of transformation efficiencies by cell type and made that information available here so that the researcher can choose the best possible method for their particular use-case. Furthermore, to create the gRNAs, Thermo Fisher offers either GeneArt CRISPR Strings DNA Fragments that can be transfected and expressed in vivo to create the gRNAs. Alternatively, they offer a gRNA synthesis kit that can transcribe short fragment dsDNAs into gRNAs in under 4 hours.
Instead of having to confirm successful CRISPR editing by genome sequencing, Thermo Fisher created the GeneArt Genomic Cleavage Selection Kit for quick and reliable functionality testing. It uses a fluorescence reporter system that only becomes active in successfully edited cells, which can then be enriched or selected through cell sorting. This methodology is particularly useful for difficult-to-transfect cells, and promises to save lots of time for researchers that might expect low cleavage activity in their CRISPR assay. Altogether, with Thermo Fisher’s CRISPR product suite, fully edited cells can be achieved in under a week from design to editing verification.
To make it even easier on researchers, Thermo Fisher has created and amassed the world’s largest collection of pre-engineered CRISPR cell lines; over 14,000 cell lines to choose from. These isogenic cell lines include the most popular knock-outs, knock-ins, and double knock-outs to quickly characterize a certain gene's role in cellular processes, disease development, or drug response.
While Thermo Fisher Scientific has been offering GeneArt Gene Synthesis, Protein Production, and Directed Evolution services for years, they continue to optimize each process and have recently launched a new initiative to offer low-cost synthetic gene libraries.
Thermo Fisher Scientific differentiates itself from other gene synthesis providers by offering a multi-parameter GeneOptimizer algorithm that includes, yet goes far beyond, the usual codon optimization most providers offer. By taking a multi-parameter approach, their GeneArt GeneOptimizer primes your gene for the highest possible in vivo expression levels. These genes can be delivered either directly cloned into a vector of your choice or as dsDNA fragments that can be cloned with any technology. They are ISO-certified, and of course all sequences are 100% verified. Thermo Fisher Scientific has recently expanded the versatility and power of their GeneArt Strings DNA Fragments by allowing library creation with up to 90 bp of randomization in fragments ranging from 200-2000 bp. Furthermore, while most gene synthesizers can only do a limited set of random nucleotides, Thermo Fisher allows you to designate mixed bases with the full IUPAC code of DNA nucleotides. While this level of customization typically takes several weeks, Thermo Fisher processes GeneArt Strings DNA Libraries in 10-15 business days.
Schematic of GeneArt Strings DNA Libraries showing the maximum 3 separate sites of 30 bp randomization.Customers have used GeneArt Gene Synthesis for numerous applications including quickly screening many enzymes for substrate affinity and expressing human proteins in E. coli. Their stories are available here, and anyone is welcome to share their own experience to be featured on the Thermo Fisher website.
Thermo Fisher Scientific offers several options for protein production depending on your needs. All custom Gene to Protein projects will go through a pilot production phase where the scientists at Thermo Fisher will ensure your protein of interest can be expressed. From there the user can decide whether they would like all of the purified protein yielded from a given volume of cell culture, or alternatively specific amount of purified protein.
Lastly, Thermo Fisher Scientific offers advanced GeneArt Directed Evolution options. Between site-directed mutagenesis, site-saturated mutagenesis, combinatorial libraries, and controlled randomization libraries they have almost every possible randomization application covered for gene variants. With this impressive suite of tools and services Thermo Fisher Scientific has proven itself a forerunner in the emerging and quickly growing synthetic biology community for genome engineering and gene synthesis. At their current pace, they are on track to become one of the most versatile resources for scientists interested in expediting their research.
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