MuA Transposase Host Range: Bacterial Species Suitable for Mutagenesis and Library Construction

Beyond the Published Mu Literature

The practical applicability of MuA transposition is likely much broader than the currently published literature illustrates.

Many researchers are familiar with the Tn5 transposome systems, which use a conceptually similar workflow: a purified transposase is assembled onto DNA in vitro to form a stable nucleoprotein complex that is subsequently introduced into cells. Commercial Tn5 technologies have been successfully applied to a wide range of Gram-negative and Gram-positive bacteria.

Because MuA transposase can likewise be delivered as a preassembled transposition complex, bacterial species that have proven amenable to Tn5 mutagenesis are excellent candidates for MuA-mediated mutagenesis as well.

While each organism should be evaluated experimentally, published results suggest that transposome-based mutagenesis can be applied to a remarkably broad spectrum of microorganisms spanning industrial, environmental, agricultural, and medically important species.

Applications of MuA Across Diverse Bacterial Species

MuA transposase is suitable for a wide variety of applications, including:

• Random insertion mutagenesis
• Genome-wide mutant library construction
• Functional genomics studies
• Gene discovery and pathway analysis
• Identification of essential genes
• Metabolic engineering
• Strain improvement
• Screening for antibiotic resistance mechanisms
• Mapping genotype-phenotype relationships
• Stable chromosomal integration of heterologous DNA
• Development of high-diversity insertion libraries

Because MuA insertions exhibit minimal sequence bias and can generate highly diverse insertion populations, the system is particularly valuable for large-scale screening applications.

Is MuA Suitable for Your Organism?

MuA transposase is often a strong candidate when any of the following apply:

• The organism can be transformed or electroporated.
• Tn5-based mutagenesis has previously been reported.
• The species belongs to a bacterial genus related to published Mu hosts.
• Large-scale mutant libraries are required.
• Stable genomic integration is desired.
• Existing genetic engineering tools are limited or unavailable.

Even if your organism has not yet been reported in the Mu literature, successful Tn5 mutagenesis is often a strong indicator that MuA-based approaches may also be feasible. Compared to wild type MuA, the hyperactive version of MuA (v.3 MuA, in vivo integrator) is ten times more efficient in its genomic integration capacity. Thus, it is always adviced to use v.3 MuA when new bacterial strains are being engineered or when highly diverse genomic insertion mutant libraries are being generated.

Bacterial/Archaeal Species Reported for MuA or Tn5-Based Mutagenesis

The table below summarizes microbial species reported in the published Mu literature and/or commonly used with Tn5-based transposome systems. Together, these examples illustrate the broad applicability of transposome-based mutagenesis across various microorganisms.

Are You Not Seeing Your Favourite Organism on the List?

The species listed above likely represent only a subset of microorganisms that have been successfully modified using Mu- or Tn5-based transposition systems.

Because MuA transposition relies on externally supplied transposition machinery, many additional bacterial species may be suitable candidates for MuA-mediated mutagenesis, insertion library construction, and genome engineering.

If your organism can be transformed or electroporated, we would be happy to discuss whether MuA transposase may be suitable for your project.

Explore Hyperactive MuA Transposases

To support efficient mutagenesis and genome engineering workflows, Domus Biotechnologies offers hyperactive MuA transposases optimized for custom transpososome assembly, mutant library construction, and chromosomal integration.

Whether you are working with a model organism or exploring a challenging non-model species, our hyperactive MuA variants provide a powerful platform for transposition-based genome engineering.

Ready to start your project? Explore our Hyperactive MuA Transposases and find the right enzyme for your application.

Explore Genomic Integration Kits

Looking for a ready-to-use solution? Our Genomic Integration Kits contain pre-assembled Mu DNA transposition complexes for efficient mutagenesis and stable genomic integration across a broad host range. Kits are available for Gram-negative bacteria, Gram-positive bacteria, and mammalian cells.

References

1. Kiljunen S, Pajunen MI, Dilks K, Storf S, Pohlschroder M, Savilahti H.: Generation of a comprehensive transposon insertion mutant library for the model archaeon Haloferax volcaniiand its use for gene discovery. 2014: BMC Biology 12:103.
2. Lamberg A, Nieminen S, Qiao M, Savilahti H.: An efficient insertion mutagenesis strategy for bacterial genomes based on the phage Mu DNA transposition reaction. 2002: Applied and Environmental Microbiology 68:705-712.
3. Rasila TS, Pajunen MI, Savilahti H.: Mu transpososome activity-profiling yields hyperactive MuA variants for genome engineering applications. 2018: Nucleic Acids Research 46:2755-2771.
4. Laasik E, Ojarand M, Pajunen MI, Savilahti H, Mäe A.: Novel mutants of Erwinia carotovoradefective in plant cell wall degrading enzyme production generated by Mu transpososome-mediated insertion mutagenesis. 2005: FEMS Microbiology Letters 243:93-99.
5. Pajunen MI, Pulliainen AT, Finne J, Savilahti H.: Generation of transposon insertion mutant libraries for Gram-positive bacteria by electroporation of phage Mu DNA transposition complexes. 2005: Microbiology 151:1205-1214.
6. Tu Quoc PH, Genevaux P, Pajunen MI, Savilahti H, Georgopoulos C, Schrenzel J, Kelley WL.: Isolation and characterization of biofilm formation-defective mutants of Staphylococcus aureus. 2007: Infection and Immunity 75:1079-1088.
7. Lanckriet A, Timbermont L, Happonen LJ, Pajunen MI, Pasmans F, Haesebrouck F, Ducatelle R, Savilahti H, Van Immerseel F.: Generation of a single-insertion mutant library in Clostridium perfringensusing a phage Mu-derived transposon. 2009: Applied and Environmental Microbiology 75:2638-2642.
8. Murooka Y, Takizawa N, Harada T.: Expansion of the host range of bacteriophage Mu. 1981: Journal of Bacteriology 145:358-368.

This paper established that phage Mu can be propagated in a number of bacterial species, showing that in these species there exists no inherent system(s) to restrict the processes involved in the life cycle of Mu. Accordingly, transpososome-mediated Mu mutagenesis is expected to be feasible in these species.

9. Epicentre Biotechnologies. EZ-Tn5™ Transposome™ Technology Guide. https://www.genetargetsolutions.com.au/wp-content/uploads/2014/06/Epicentre-Mutagenesis-Tn5-Transposomics.pdf

This document lists a number of organisms, in which Tn5 transpososome-mediated mutagenesis has been reported to be feasible. Unfortunately, the document lacks references for the verification of the disclosed results. However, published Tn5 transpososome delivery data and references for various organisms can be found e.g. from the Web of Science or Google Scholar.

10. Wu Z, Xuanyuan Z, Li R, Jiang D, Li C, Xu H, Bai Y, Zhang X, Turakainen H, Saris PEJ, Savilahti H, Qiao M.: Mu transposition complex mutagenesis in Lactococcus lactis: identification of genes affecting nisin production. 2009: Journal of Applied Microbiology 106:41-48.
11. Paatero AO, Turakainen H, Happonen LJ, Olsson C, Palomäki T, Pajunen MI, Meng X, Otonkoski T, Tuuri T, Berry C, Malani N, Frilander MJ, Bushman FD, Savilahti H.: Bacteriophage Mu Integration in Yeast and Mammalian Genomes. 2008: Nucleic Acids Ressearch 36:e148.
12. Bai F, Li YL, Xu HJ, Xia HM, Yin TF, Yao HM, Zhang L, Zhang XM, Bai YL, Jin SG, Qiao M. Identification and functional characterization of pfm, a novel gene involved in swimming motility of Pseudomonas aeruginosa. Gene 2007;401, 19-27.
13. Rantakokko A, University of Turku, Finland Personal communication. 2026; unpublished results with the use of v.3 MuA (In vivo Integrator by Domus Biotechnologies).
14. Weber C, University of Georgia, USA. Personal communication. 2026; unpublished results with the use of v.3 MuA (In Vivo Integrator by Domus Biotechnologies).