Mutation, selection, reproduction, repeat for millions of generations—every biologist is familiar with how evolution has resulted in the incredible diversity and adaptations found in the natural world. Directed evolution uses a similar strategy to create biomolecules for use in diverse fields spanning industrial production, medical science, and basic research. The crucial difference is that, with directed evolution, results can be achieved much more quickly, in many cases with just a few rounds of mutagenesis and selection.

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Figure 1: Directed evolution

In vitro molecular evolution strategies (Figure 1) are widely regarded as the most efficient methods for creating proteins and regulatory sequences with improved or novel properties. Life Technologies GeneArt® Directed Evolution services cover a variety of different approaches for creating genetic variants. Paired with appropriate screening methods, our directed evolution technologies can create the genetic diversity you need to obtain proteins with the characteristics you need.

Synthetic Variant Libraries Help You Find Your Protein Faster

Conventional mutagenesis protocols rely on virtually random mutations. For example, with error-prone PCR, the position and nature of the mutations cannot be controlled. As a result, mutant libraries contain many silent mutations and ill-placed stop codons. Perhaps of greater concern, however, is that only a tiny fraction of all possible sequence variants can be generated, so finding the best possible variant is extremely unlikely. See an example.

DNA shuffling can be a powerful approach for creating diversity, but is subject to drawbacks including the requirement for stretches of homologous sequence to serve as sites for recombination at a reasonably high rate, and the need to obtain DNA as starting material. In addition traditional DNA shuffling typically does not separate and recombine adjacent single-nucleotide polymorphisms—changes which have the potential to result in useful protein variants.

Major benefits of GeneArt® Directed Evolution Services

  • Most service offerings do not require DNA template—DNA is synthesized from your sequence information
  • Complete control over every important feature of the library—Create rational diversity where it is likely to have the most impact
  • Significantly reduced screening effort compared to conventional mutagenesis methods
  • Generate variants that cannot be created using conventional methods
  • Maximum sequence integrity in unmutated regions, maximum variation where you want it, and maximum insert size
  • Improved likelihood of obtaining useful variants
GeneArt® synthetic libraries overcome many of the limitations of conventional variant-library construction techniques. De novo gene synthesis enables construction of virtually any gene variation so that your library encodes maximum variability. Supported by GeneArt® GeneOptimizer® assisted sequence design, synthetic libraries achieve thorough representation of desired variants with the specified distribution of nucleotides in areas targeted for partial degeneration or full randomization. The GeneArt® gene synthesis process simultaneously minimizes the introduction of unwanted mutants and erroneous changes to non-mutated portions of your constructs. This approach dramatically reduces the number of variants—economizing screening time, reagents, and effort—while increasing your chance of success. Because library synthesis is largely automated, we can offer rapid production times so that you can get started quickly. Our rigorous quality control systems include sequencing, statistical sequence analysis, and real-time PCR diversity analysis (control procedures are tailored to individual product lines).

Tell us about your project for more information and to receive information on pricing and production time.

GeneArt® Directed Evolution Services

GeneArt® Combinatorial Libraries

New! Maximum diversity via TRIM technology

Completely synthetic process for introducing random variation in multiple codons (up to 1012 variants) with complete customization of the amino acid composition. Ideal for introducing targeted variation while maintaining maximum framework integrity.

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GeneArt® Mutagenesis Service

Site-directed mutagenesis

Rapid, economical mutagenesis of existing DNA templates using a PCR-based approach.

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GeneArt® Site-Saturation Mutagenesis

Systematic mutagenesis

Systematic mutagenesis to substitute the wild type codon at specific positions with codons for as many as all 19 non wild type amino acids. Many options available to fit your precise needs.

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GeneArt® Controlled Randomization Service

Random substitutions

Advanced technology for introducing unbiased random mutations at the frequency you request, in the regions of the gene you specify. Mutatagenize the entire ORF or confine variation to selected regions (up to 1011 variants).

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GeneArt® Truncation Libraries

Truncation without frameshifts

Customer defined truncations, including simultaneous 5’ and 3’ deletions, without inducing out-of-frame mutations.

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