mRNA-sequencing whole transcriptome analysis of a single cell on the SOLiD system.
We have developed a sequencing-based gene expression profiling assay at single-cell resolution by combining a modified single-cell whole transcriptome amplification method with the next generation sequencing technique, the SOLiD system. Using this assay, we have shown that blastomeres in a four-cell stage embryo have similar gene expression, which is compatible with the fact that they have similar developmental potential. We proved that compared with cDNA microarray technique, our single-cell cDNA SOLiD sequencing assay can detect expression of thousands of more genes. Moreover, for the genes detected by microarray and SOLiD sequencing, our assay detected new transcript variants for a large proportion of them, which confirms unambiguously at single-cell resolution that the transcriptome complexity is higher than expected traditionally. Finally, by using our assay to Dicer knockout (KO) and Ago2 KO oocytes, we showed that a significant amount of transposons were up-regulated abnormally in Dicer/Ago2 KO mature oocytes compared with wild-type controls.
Lao
Kai Q
KQ
Applied Biosystems, a division of Life Technologies Corporation, Genetic Analysis Business Unit, Foster City, California 94404, USA. kai.lao@lifetech.com
Tang
Fuchou
F
Barbacioru
Catalin
C
Wang
Yangzhou
Y
Nordman
Ellen
E
Lee
Clarence
C
Xu
Nanlan
N
Wang
Xiaohui
X
Tuch
Brain
B
Bodeau
John
J
Siddiqui
Asim
A
Surani
M Azim
MA
D000818:Animals; D002874:Chromosome Mapping; D018076:DNA, Complementary / Q000378:metabolism; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005821:Genetic Techniques; D051379:Mice; D018345:Mice, Knockout; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D009865:Oocytes / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA / Q000379:methods*
D018076:DNA, Complementary; D012333:RNA, Messenger