Methods and compositions for translational profiling and molecular phenotyping
Methods and compositions are provided for translational profiling and molecular phenotyping of specific tissues, cells and cell subtypes of interest. The methods provided herein facilitate the analysis of gene expression in the selected subset present within a heterogeneous sample.
1. A nucleic acid molecule comprising a nucleotide sequence encoding a ribosomal fusion protein comprising mouse ribosomal protein L10a having the amino acid sequence of SEQ ID NO: 30 fused to an optically detectable protein, wherein the nucleotide sequence is operably linked to a mouse endogenous regulatory sequence that causes expression of the fusion protein in a chosen cell type, and wherein the ribosomal fusion protein is incorporated in an intact ribosome that translates and/or binds mRNA.
2. The nucleic acid molecule of claim 1 , wherein the optically detectable protein is a fluorescent protein.
3. The nucleic acid molecule of claim 2 , wherein the fluorescent protein has greater than approximately 200 amino acids.
4. The nucleic acid molecule of claim 3 , wherein the fluorescent protein is an enhanced green fluorescent protein (eGFP).
5. The nucleic acid molecule of claim 1 , wherein the cell type is neuronal.
6. The nucleic acid molecule of claim 1 , wherein the cell type is located in the central nervous system.
7. The nucleic acid molecule of claim 6 , wherein the cell type is an astrocyte.
8. The nucleic acid molecule of claim 6 , wherein the cell type is a cholinergic neuron.
9. The nucleic acid molecule of claim 6 , wherein the location of the cell type is selected from the group consisting of spinal cord, cerebellum, hypothalamus, substantia nigra, ventral tegmental area, and striatum.
10. A mouse comprising a nucleic acid molecule comprising:
a nucleotide sequence encoding a ribosomal fusion protein comprising mouse ribosomal protein L10a having the amino acid sequence of SEQ ID NO: 30 fused to an optically detectable protein, wherein the nucleotide sequence is operably linked to an endogenous mouse regulatory sequence that causes expression of the fusion protein in a chosen cell type, and wherein the ribosomal fusion protein is incorporated in an intact ribosome that translates and/or binds mRNA.
11. The mouse of claim 10 , wherein the optically detectable protein is a fluorescent protein.
12. The mouse of claim 11 , wherein the fluorescent protein has greater than approximately 200 amino acids.
13. The mouse of claim 12 , wherein the fluorescent protein is an enhanced green fluorescent protein (eGFP).
14. The mouse of claim 10 , wherein the cell type is neuronal.
15. The mouse of claim 10 , wherein the cell type is located in the central nervous system.
16. The mouse of claim 15 , wherein the cell type is an astrocyte.
17. The mouse of claim 15 , wherein the cell type is a cholinergic neuron.
18. The mouse of claim 15 , wherein the location of the cell type is selected from the group consisting of spinal cord, cerebellum, hypothalamus, substantia nigra, ventral tegmental area, and striatum.