Identification of miRNAs in a liver of a human fetus by a modified method.
BACKGROUND
miRNAs are 17-25 nucleotides long RNA molecules that have been found to regulate gene expression in human cells. There are studies showing that different groups of miRNAs are involved in development of different tissues. In hepatocytes there are reported particular types of miRNAs that regulate gene expression.


METHODS
We established a human fetal liver cDNA library by a modified cloning protocol. Then plasmid isolation from the colonies was performed. After sequencing and database searching, the miRNAs were recognized. RT-PCR and sequencing were carried out to validate the miRNAs detected. Real-time PCR was used to analyze the expression of each miRNA.


RESULTS
One novel miRNA was discovered, together with another 35 previously-known miRNAs in the fetal liver. Some of them existed in variants. The miRNAs identified were validated by RT-PCR and sequencing. Quantitative analysis showed that they have variable expression.


CONCLUSIONS
Our results indicate that a special group of miRNAs may play an important role in fetal liver development in a synergistic manner.
Liu
Dong
D
Department of Medical Genetics, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Fan
Jing
J
Mei
Manxue
M
Ingvarsson
Sigurdur
S
Chen
Huiping
H
D001483:Base Sequence; D002478:Cells, Cultured; D005260:Female; D005333:Fetus / Q000378:metabolism; D015723:Gene Library; D005821:Genetic Techniques; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000196:embryology* / Q000378:metabolism*; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid
D035683:MicroRNAs