Bacterial Protein Injection for Stem Cell Differentiation and Genome Editing

Technology #15695

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Researchers
Shouguang Jin Ph.D.
Naohiro Terada M.D.
Fang Bai PhD
Managed By
April Kilburn
Assistant Director 352-392-8929

Safely and Efficiently Differentiates and Edits Genomes of Stem Cells

This bacterial injection of proteins can deliver specific transcription factors to achieve directed differentiation of pluripotent stem cells. Furthermore, it can also inject TALEN proteins for genome editing purpose. Stem cells are currently employed for various research measures, including disease studies, regenerative medicine, cancer therapy, and drug safety testing. Induced pluripotent stem cells (iPSCs) are cells taken from any tissue and modified to behave like embryonic stem cells, with the ability to form all adult cell types. Their versatility provides a wealth of knowledge regarding cell science and molecular biology, showing great potential for disease treatment and prevention. Researchers at the University of Florida have discovered a safe method of protein delivery for iPSC differentiation and genome editing, which is low in cost, high in efficiency, and readily scalable. This bacterial injection has been successfully applied to the differentiation of pluripotent stem cells into cardiac muscle cells and genome editing of the pluripotent stem cells.

Application

Directed differentiation and genome editing of pluripotent stem cells as well as primary cell types.

Advantages

  • Directly injected into target cells using a non-virulent delivery strain, eliminating safety concerns associated with the use of viral or DNA delivery
  • Capable of injecting proteins into various mammalian cell types, including primary (differentiated as well as stem cells) or transformed cell lines, offering a wide range of target cell types
  • Can translocate various types of proteins, including transcriptional factors that can target to nucleus and trigger downstream gene expression
  • Has no need for protein purification, making it easily adaptable to large scale production

Technology

This bacterial protein injection system uses highly efficient protein delivery machinery, called Type III Secretion System (T3SS), a needle-like surface structure encoded by Pseudomonas aeruginosa designed to inject bacterial proteinous toxins directly into the cytoplasmic compartment of host cells, without the need for bacteria to get inside the host cells. This naturally occurring high efficiency protein injection machinery can deliver proteins into mammalian host cells. By fusing a short N-terminal secretion sequence (54AA peptide) with a non-cytotoxic P. aeruginosa strain for T3SS-dependent injection, University of Florida researchers are able to deliver various target proteins into mammalian cells, including transcriptional factors that can turn on the expression of their target genes and TALEN nucleases that introduce double-stranded cleavage on intended target sites.