siRNA in primary lymphocyte B - Which reagent do you recomend? (Feb/06/2008 )

Hi everybody,

I would like some advise for tranfecting siRNA in primary B-lymphocyte of mouse spleen?

Which are the reagents that keep more viability? and which are their efficiency?

thanks in advance

I have also very similar question " siRNA delivery in human B cell lines" and still waiting for suggestions. I was advised to use “Retrovirus vector”.

If you would consider doing your knockdowns with antisense, then the combinations of Morpholino antisense oligos with either electroporation or Endo-Porter endosomal release agent have worked well in lymphocytes.

Here are a few references describing delivery of Morpholinos into Jurkat cells by electroporation:

Combs J, Kim SJ, Tan S, Ligon LA, Holzbaur EL, Kuhn J, Poenie M. Recruitment of dynein to the Jurkat immunological synapse. Proc Natl Acad Sci U S A. 2006 Oct 3;103(40):14883-8. Epub 2006 Sep 21.

Matter N, Konig H. Targeted 'knockdown' of spliceosome function in mammalian cells. Nucleic Acids Res. 2005 Feb 24;33(4):e41.

So far, I have not seen a published report of delivery into lymphocytes by Endo-Porter, but we have heard from some Morpholino users about successful Morpholino delivery into lymphocytes by Endo-Porter. For Endo-Porter references, see the bottom of this page: http://www.gene-tools.com/node/24

For a searchable database of over 2000 Morpholino publications, see pubs.gene-tools.com