I am interested in a gene which is null lethal but I need to temporary induce diminished capacity. If a cell is homozygous is it possible to induce heterozygous phenotypes or a partial knockout from WT with siRNA or other molecular methods without virus vectors? I'm not interested in a medillian solution but an inducible molecular way. Thank you

I would like to generate a heterozygous Sod2+/- like they do in this paper in mice and possibly human: http://physiolgenomics.physiology.org/content/16/1/29.full.pdf+html

  • $\begingroup$ It would help if you gave more information about the system that you are working with - what type of cells? $\endgroup$ – Alan Boyd Nov 2 '12 at 7:04
  • $\begingroup$ @AlanBoyd im interested in mice and humans and i added the gene of interest as well. $\endgroup$ – user1357 Nov 2 '12 at 7:16
  • $\begingroup$ The tetracycline-regulated systems developed by Clontech would allow you to do something like this - the level of gene expression can be titrated by varying the concentration of tetracycline/doxycycline in the growth medium. See clontech.com/GB/Products/Inducible_Systems/… for links to more details - the early generation products might be a good place to start reading. $\endgroup$ – Alan Boyd Nov 2 '12 at 18:23
  • $\begingroup$ @AlanBoyd my growth\life medium would prohibit the use of tetracycline but thanks for using the key word titrated $\endgroup$ – user1357 Nov 3 '12 at 0:10
  • $\begingroup$ what medium is this? $\endgroup$ – Alan Boyd Nov 3 '12 at 7:01

Antisense, promoter replacement and when possible tetracycline allows one to titrate of gene expression http://www.springerprotocols.com/Abstract/doi/10.1007/978-1-59745-321-9_20#

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