FIN-Seq: transcriptional profiling of specific cell types from frozen archived tissue of the human central nervous system

被引:12
作者
Amamoto, Ryoji [1 ,2 ]
Zuccaro, Emanuela [1 ,4 ]
Curry, Nathan C. [1 ]
Khurana, Sonia [1 ]
Chen, Hsu-Hsin [1 ]
Cepko, Constance L. [2 ]
Arlotta, Paola [1 ,3 ]
机构
[1] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA
[2] Harvard Med Sch, Howard Hughes Med Inst, Blavatnik Inst, Dept Genet & Ophthalmol, Boston, MA 02115 USA
[3] Broad Inst MIT & Harvard, Stanley Ctr Psychiat Res, Cambridge, MA 02142 USA
[4] Univ Padua, Dept Biomed Sci, I-35131 Padua, Italy
关键词
NUCLEUS RNA-SEQ; SUBSTANTIA-NIGRA; GENE-EXPRESSION; NEURONAL SUBTYPES; HIGH-THROUGHPUT; DISEASE; DIVERSITY; DYNAMICS;
D O I
10.1093/nar/gkz968
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Thousands of frozen, archived tissue samples from the human central nervous system (CNS) are currently available in brain banks. As recent developments in RNA sequencing technologies are beginning to elucidate the cellular diversity present within the human CNS, it is becoming clear that an understanding of this diversity would greatly benefit from deeper transcriptional analyses. Single cell and single nucleus RNA profiling provide one avenue to decipher this heterogeneity. An alternative, complementary approach is to profile isolated, predefined cell types and use methods that can be applied to many archived human tissue samples that have been stored long-term. Here, we developed FIN-Seq (Frozen Immunolabeled Nuclei Sequencing), a method that accomplishes these goals. FIN-Seq uses immunohistochemical isolation of nuclei of specific cell types from frozen human tissue, followed by bulk RNA-Sequencing. We applied this method to frozen postmortem samples of human cerebral cortex and retina and were able to identify transcripts, including low abundance transcripts, in specific cell types.
引用
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页数:12
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