Mutation, selection, reproduction—repeated 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 is a protein engineering method that involves a few rounds of iterative variant library creation and selection for desired traits. This is a powerful tool for researchers to develop proteins with enhanced or novel functions in a short period of time.

Benefits of designing variant libraries with GeneArt services

Invitrogen GeneArt Custom DNA Libraries cover a variety of different approaches for creating genetic variants with control and efficiency. Starting from a single-base pair exchange to full synthetic libraries, we offer custom solutions to create genetic diversity and proteins with the characteristics you need.

  • No physical template required
  • Generate variants that cannot be created using conventional methods
  • Maximum variation where you want it, and maximized sequence integrity in unmutated regions
  • Helps improve the likelihood of obtaining useful variants
  • Helps significantly reduce screening efforts compared to conventional mutagenesis methods
  • Include next-generation sequencing quality control for your library (optional)


Featured GeneArt directed evolution services

Choosing a custom library

Use the decision tree (Figure 1) to choose the right GeneArt mutagenesis and library synthesis services.

Flowchart guiding selection of GeneArt mutagenesis and library synthesis services
Figure 1. Selection guide: GeneArt mutagenesis and library synthesis services.
Table 1. Comparison of the GeneArt mutagenesis and library synthesis services.
 GeneArt Strings DNA librariesGeneArt site-saturation mutagenesisGeneArt controlled randomization serviceGeneArt combinatorial libraries

Advantage

  • Cost efficient
  • Fast
  • Fewer variants to screen
  • Fast service
  • Cost efficient
  • Precise average mutation rate per construct
  • Fast service
  • High diversity
  • Control of amino acid distribution
  • No unwanted stop codons
Design flexibility+
Full IUPAC code available; however limited control over occurring amino acids		++
Systematic identification of beneficial amino acid substitutions		++
Random nucleotides within a defined region will be mutated at a defined average mutation rate		+++
TRIM technology allows for the accurate determination of amino acid ratios at each position
Correctness+
Gene synthesis process used, more unintended mutations occur		+++
Error-free template enables best possible correctness of results		++
Low ancillary mutation rate, fewer silent mutations		+++
Error-free template enables superior correctness of results
Complexity<101116–19 non-wild type amino acids per codon position>1010–1011>1011
>1012 optional
Cost$$$$$$–$$$
Production time+++
10–15 business days		++++
4–6 weeks
Quality control
  • Pool sequenced
  • Size selection
  • Pool sequenced
  • Sequencing of a sampling of individual transformants
  • Pool sequenced
  • Sequencing of a sampling of individual transformants
  • NGS option available
  • Pool sequenced
  • Sequencing of a sampling of individual transformants
  • NGS option available


Synthetic variant libraries

Synthetic variant libraries are designed to overcome some of the limitations associated with conventional mutagenesis and DNA shuffling. These libraries use advanced techniques to create a more focused and comprehensive collection of genetic variants. By precisely controlling the introduction of mutations, variant libraries can systematically explore a wider range of sequence space. This targeted approach increases the likelihood of identifying beneficial mutations and reduces the presence of non-functional variants. Additionally, variant libraries can be tailored to include specific types of mutations, such as those affecting protein function or stability, thereby enhancing the efficiency and effectiveness of the screening process.

Drawbacks of conventional mutagenesis

  • Position and nature of mutations cannot be controlled
  • Mutant libraries contain many silent mutations and ill-placed stop codons
  • Only a tiny fraction of all possible sequence variants can be generated
  • Finding the best possible variant is extremely unlikely

Drawbacks of DNA shuffling

  • Requires stretches of homologous sequences for high-efficiency recombination sites
  • Needs DNA as starting material
  • Traditional DNA shuffling typically does not separate and recombine adjacent single-nucleotide polymorphisms
  • Single-nucleotide polymorphisms have the potential to result in useful protein variants

Why choose GeneArt custom variant libraries for directed evolution

GeneArt DNA libraries for protein engineering address many of the challenges associated with 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 the GeneArt GeneOptimizer algorithm for sequence design, synthetic libraries achieve thorough representation of desired variants with the specified distribution of nucleotides in areas targeted for partial degeneracy or full randomization. The GeneArt Gene Synthesis process simultaneously minimizes the introduction of unwanted mutants and erroneous changes to unmutated portions of your constructs. This approach dramatically reduces the number of variants—economizing screening time, reagents, and effort—while helping to increase 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 (and optional next-generation sequencing), statistical sequence analysis, and real-time PCR diversity analysis (control procedures are tailored to individual product lines).

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

Search

  • Contact Us
  • Quick Order