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Getting Morpholinos into cultured cells

There are three usual options to consider for cell culture experiments with Morpholinos. There are other methods, but most labs choose one of these.

(1) If your cells can tolerate electroporation, that is an inexpensive option for delivery of standard Morpholino oligos. However, electroporation causes high cell mortality for many cell types.

(2) Use Endo-Porter to deliver a standard Morpholino oligo. Endo-Porter is most effective in its DMSO formulation. The Endo-Porter peptide is poorly soluble in water and when the DMSO formulation is pipetted into culture medium, it is important to immediately swirl the medium to disperse the Endo-Porter. The Endo-Porter peptide will form complexes in the aqueous medium and swirling keeps these complexes small and abundant; these small and abundant complexes produce the highest delivery efficacy. The Endo-Porter aqueous formulation is provided because some undifferentiated cell types are triggered to differentiate by small concentrations of DMSO. The aqueous formulation is about a quarter as effective as the DMSO formulation and should only be used if needed. To treat around 150 ml of culture medium, you will need one order of Endo-Porter ($200) and 300 nanomoles of standard Morpholino ($400). The 150 ml estimate can vary, as different cell lines have different optimal concentrations of Endo-Porter and the required dose of a Morpholino oligo will vary with the sequence; standard Morpholinos are typically added to a final concentration of 2 to 10 micromolar in the cell medium.

(3) Vivo-Morpholinos can be used in cell cultures by adding the Vivo-Morpholino to the medium on the cells, with no other delivery system required. Vivo-Morpholinos are typically added to a final concentration of 2 to 10 micromolar in the cell medium. Vivo-Morpholinos work with most cell types tested but some are sensitive to the Vivo-Morpholino's attached delivery moiety, an octaguanidinium dendrimer. 400 nanomoles of a Vivo-Morpholino costs $700, more than the combination of a standard Morpholino with Endo-Porter but you receive more oligo.

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