Domus v.3 MuA Transposase (in vivo Integrator) is a hyperactive MuA transposase variant (1). It has a superior performance especially in genomic integration assays, both with bacteria and eukaryotic cells. When v.3 MuA Transposase is incubated in vitro with Mu transposon in the absence of Mg2+, stable transpososomes are formed. These complexes can be electroporated into living cells, where they encounter Mg2+ and become activated for transposition. Subsequently, transpososomes are able to integrate the delivered transposon DNA into the host chromosome. Transposons can contain any DNA (e.g. selectable markers or other genetic elements), between MuA binding sites at both ends.
v.3 MuA Transposase (in vivo Integrator)
- Preparation of Mu transpososomes for efficient gene delivery following electrotransformation
- Highly efficient gene delivery for a variety of bacteria, and also for yeast and mammalian cells (1-5)
- Can be used to generate exhaustive insertion mutant libraries for many types of micro-organisms, also to archaeal species (6)
- Nearly random insertion profile (7,8)
Scheme for genomic integration. A tetramer of MuA transposase and Mu transposon ends assemble into stable transpososomes in vitro. Following electroporation, the transpososomes encounter Mg2+ ions in vivo and integrate transposon DNA into the chromosome.
1. Rasila,T.S., Pulkkinen,E., Kiljunen,S., Haapa-Paananen,S., Pajunen,M.I., Salminen,A., Paulin,L., Vihinen,M., Rice,P.A. and Savilahti,H. (2018) Mu transpososome activity-profiling yields hyperactive MuA variants for highly efficient genetic and genome engineering. Nucleic Acids Res., 46, 4649–4661.
2. Lamberg A, Nieminen S, Qiao M, Savilahti H (2002) Efficient insertion mutagenesis strategy for bacterial genomes involving electroporation of in vitro-assembled DNA transposition complexes of bacteriophage Mu. Appl Environ Microbiol 68:705-712
3. Pajunen MI, Pulliainen AT, Finne J, Savilahti H (2005) Generation of transposon insertion mutant libraries for Gram-positive bacteria by electroporation of phage Mu DNA transposition complexes. Microbiology 151:1209-1218
4. 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 (2008) Bacteriophage Mu integration in yeast and mammalian genomes. Nucleic Acids Res 36:e148
5. Tu Quoc PH, Genevaux P, Pajunen M, Savilahti H, Georgopoulos C, Schrenzel J, Kelley WL (2007) Isolation and characterization of biofilm formation-defective mutants of Staphylococcus aureus. Infect Immun 75:1079-1088
6. Wu Z, Xuanyuan Z, Li R, Jiang D, Li C, Xu H, Bai Y, Zhang X, Turakainen H, Saris PE, Savilahti H, Qiao M (2009) Mu transposition complex mutagenesis in Lactococcus lactis-identification of genes affecting nisin production. J Appl Microbiol 106:41-8
7. Kiljunen S, Pajunen MI, Dilks K, Storf S, Pohlschroder M, Savilahti H (2014) Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii , and its use for gene discovery. BMC Biol 12:103
8. Haapa-Paananen S, Rita H, Savilahti H (2002) DNA transposition of bacteriophage Mu. A quantitative analysis of target site selection in vitro. The Journal of Biological Chemistry 277:2843-2851
9. Mizuuchi M, Mizuuchi K (1993) Target site selection in transposition of phage Mu. Cold Spring Harb Symp Quant Biol 58:515-523
v.3 MuA Transposase (in vivo Integrator) is covered by International Patent No. WO 2014/013127 Al
For Research Use Only. Not for use in diagnostic procedures.