Bio-Techne has been granted a patent for TcBuster transposases and transposons, along with their systems and methods of use. The patent specifically covers a mutant TcBuster transposase with specific amino acid substitutions. GlobalData’s report on Bio-Techne gives a 360-degree view of the company including its patenting strategy. Buy the report here.
According to GlobalData’s company profile on Bio-Techne, Peptide pharmacophores was a key innovation area identified from patents. Bio-Techne's grant share as of September 2023 was 40%. Grant share is based on the ratio of number of grants to total number of patents.
Mutant tcbuster transposase with specific amino acid substitutions
A recently granted patent (Publication Number: US11760983B2) discloses a mutant TcBuster transposase that has improved transposition efficiency compared to the wild-type transposase. The mutant transposase has specific amino acid substitutions (E247K, L268T, and Y284I) and is at least 70% identical to the full-length sequence (SEQ ID NO: 1). Additionally, the mutant transposase can have additional amino acid substitutions (N85S, D99A, N209E, T219S, V356L, V377T, E469K, C470M, A472P, K490I, C512E, or any combination thereof) or (D189A, V377T, E469K, K573E, E578L, or any combination thereof) for further enhancement.
The patent also describes a fusion transposase that combines the TcBuster transposase sequence with a DNA sequence specific binding domain and/or an additional Nuclear Localization Signal sequence. The fusion transposase can have the same amino acid substitutions as the mutant transposase and exhibits increased transposition efficiency. The DNA sequence specific binding domain can be a TALE domain, zinc finger domain, AAV Rep DNA-binding domain, or a combination thereof. The fusion transposase and the DNA binding domains and/or Nuclear Localization Signal sequences can be separated by one or more linkers.
The patent further covers polynucleotides encoding the mutant transposase and the fusion transposase, which can be used for genome editing. The polynucleotides can be DNA, messenger RNA (mRNA), or chemically modified mRNA, and can be present in DNA vectors, minicircle plasmids, or other nucleic acid sequences encoding a transposon recognizable by the mutant or fusion transposase. The polynucleotides can also be codon-optimized for expression in human cells.
Methods of genome editing using the mutant transposase or the polynucleotide encoding it are also described in the patent. The methods involve introducing the mutant transposase or the polynucleotide into cells, along with a transposon recognizable by the mutant transposase. Various techniques such as electroporation, microinjection, calcium phosphate precipitation, and liposome-mediated transfection can be used for introducing the transposase and transposon into cells. The cells can be primary cells, including immune cells.
The patent also discloses a system for genome editing that includes the mutant transposase and a transposon recognizable by it. The transposon can have a cargo cassette positioned between two inverted repeats, with the left and right inverted repeats having specific nucleic acid sequences. The cargo cassette can be in a reverse direction.
Overall, this patent provides valuable insights into the development of mutant TcBuster transposases and fusion transposases for efficient genome editing applications.