Part:BBa_K4869001

Super PiggyBac Transposase Expression Vector

General Information & Literature Review:

The PiggyBac (PB) transposon is a type of mobile genetic element capable of transferring genetic material between vectors and chromosomes in a highly efficient manner via a "cut and paste" mechanism. The PB transposase identifies specific inverted terminal repeat sequences (ITRs) located at both ends of the transposon vector during the transposition process and moves the contents from their original sites, integrating them efficiently into TTAA chromosomal sites. This piggyBac transposon system makes it easy to transfer genes of interest between the two ITRs in the PB vector into target genomes.

The super piggyBac transposase expression vector also has a circular shape and consists of 6,964 base pairs, making it relatively smaller than the BII-CMV-AfmW3A vector. This vector contains CMV promoter, SV40 late 16s intron, Super PiggyBac Transposase, and SV40 poly-A. PiggyBac is a transposon, a DNA sequence that can move around within the genome of a cell. It has been adapted for use in genetic engineering, allowing the insertion, deletion, or movement of genes within the genome of various organisms. It is a hyperactive version of the enzyme that efficiently integrates any-sized DNA insert into the genome. The Super PiggyBac transposase expression vector features a strong CMV promoter to provide robust transposase expression. The PiggyBac transposase coding sequence has been optimized for high expression, stability, and integration activity in mammalian cells.

Application in Our Project:

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Figure: Super PiggyBac Transposase Expression Vector contains Super PiggyBac Transposase gene (vector sequence).

Super piggyBac transposase expression vector is a plasmid vector that was used to supplement the process of BII-CMV-AfmW3A vector genes successfully getting inserted into the host genome (HEK293). The Super piggyBac system is a powerful tool for introducing and stably integrating genetic material into host cells, ensuring long-term and stable expression of the desired genes. With the BII-CMV-AfmW3A vector, the super piggyBac transposase expression vector was used in vector transfection, which is a process of artificially putting external DNA into the target cell. This vector transfection facilitated the engineering process of HEK293 cells to produce the Afamin-Wnt3a protein complex by the insertion of Afamin and Wnt3a genes into the host cell.

References:

Biosciences, S. (n.d.). PiggyBacTM Transposon Vector System. systembio. www.systembio.com/wp/wp-content/uploads/Manual_PiggyBac_System.pdf.

Singh A. & Goodwin M. PiggyBac-ing Through the Genome Editing Field. blog.addgene.org/piggybac-ing-through-the-genome-editing-field.

Woodard LE, Wilson MH. piggyBac-ing models and new therapeutic strategies. Trends Biotechnol. 2015 Sep;33(9):525-33. doi: 10.1016/j.tibtech.2015.06.009. Epub 2015 Jul 23. PMID: 26211958; PMCID: PMC4663986.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]