14-3-3

Dr. Yi Zhou's Thy1-R18 Transgenic Mice

See SFN abstracts (2010) 53.1/M17 and (2011) 790.09/FF16

From Kourtney Graham's Honor's Thesis:

In order to study the role of 14-3-3 proteins in the nervous system_ in vivo_, we have successfully created lines of neuronal-specific transgenic mice that may inhibit 14-3-3 dependent cellular processes. These transgenic mice express a YFP-fused peptide (R18) which binds to endogenous 14-3-3, and thus blocking its capability to interact with its binding partners; and are therefore considered to be a functional knockout. The transgenes are expressed under the control of the Thy-1 promoter which is neuronal-specific and produces variation of transgene expression in different brain regions, which is great among founder lines but minimal among descendants of a single founder (Feng et al. 2000). To minimize strain-dependent genetic variation, we have backcrossed the founder lines with wild-type C57BL/6 mice for at least 8 generations.

From Sun et al PMID 23861400 : Glutamatergic kainate receptors (KAR) KAR-EPSCs at mossy fiber-CA3 synapses decay significantly faster in the 14-3-3 functional knock-out mice (but no difference in NMDA-R or AMPA-R EPSC in same synapses)

R18 peptide paper: PMID 10493820

Thy-1 Promoter paper: PMID 11086982

YFP is 239 amino acids long (717 bp) according to plasmid map of Invitrogen Vivid ColorsTM pcDNATM/6.2 N-terminal EmGFP/YFP-DEST vectors see p, 28 of the manual. R18 is 21 amino acids long (63 bp), so total YFP-R18 fusion protein should be ~ 260 amino acids (780 bp).

Vogt, Peggy, "Exploring the Function of 14-3-3 Proteins in Neurodegenerative Disease" (2011). Honors Theses. Paper 42.

C. Sun, H. Qiao, Q. Zhou, Y. Wang, Y. Wu, Y. Zhou, et al., Modulation of GluK2a Subunit-containing Kainate Receptors by 14-3-3 Proteins, J Biol Chem. 288 (2013) 24676–24690. PMID 23861400