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Dev Dyn. December 1, 2012; 241 (12): 1960-72.

Cell type-specific translational profiling in the Xenopus laevis retina.

Watson FL , Mills EA , Wang X , Guo C , Chen DF , Marsh-Armstrong N .

Translating Ribosome Affinity Purification (TRAP), a method recently developed to generate cell type-specific translational profiles, relies on creating transgenic lines of animals in which a tagged ribosomal protein is placed under regulatory control of a cell type-specific promoter. An antibody is then used to affinity purify the tagged ribosomes so that cell type-specific mRNAs can be isolated from whole tissue lysates. Here, cell type-specific transgenic lines were generated to enable TRAP studies for retinal ganglion cells and rod photoreceptors in the Xenopus laevis retina. Using real time quantitative PCR for assessing expression levels of cell type-specific mRNAs, the TRAP method was shown to selectively isolate mRNAs expressed in the targeted cell and was efficient at purifying mRNAs expressed at both high and low levels. Statistical measures used to distinguish cell type-specific RNAs from low level background and non-specific RNAs showed TRAP to be highly effective in Xenopus. TRAP can be used to purify mRNAs expressed in rod photoreceptors and retinal ganglion cells in X. laevis. The generated transgenic lines will enable numerous studies into the development, disease, and injury of the X. laevis retina.

PubMed ID: 23074098
PMC ID: PMC4536902
Article link: Dev Dyn.
Grant support: 1R01EY019960 NEI NIH HHS , R01 EY019960 NEI NIH HHS

Genes referenced: barhl2 crx fabp7 gnat1 isl2 notch1 pde6a rbpms rbpms2 rho rho.2 rpe rpl10a

Antibodies referenced: Sncg Ab1

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Agathocleous, 2009, Pubmed, Xenbase[+]

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