Screening for trans-acting factors and other factors involved in the activating or silencing of the gamma-globin gene during human ontogeny.
Researchers hope to increase gamma-globin expression by controlling potential trans-acting factors that specifically activate the gamma-globin gene in fetuses or silence this gene in adults to potentially treat sickle cell disease and beta-thalassemias. To characterize genes encoding such factors, we analyzed the differential expression of mRNAs in erythroid induction cultures of CD34+ cells derived from normal adult bone marrow, umbilical cord blood, and bone marrow from a patient with heterocellular hereditary persistence of fetal hemoglobin. Using differential-display - reverse-transcription PCR analysis, we identified a number of genes with differential expression in the above-mentioned cells. The differential expression of some genes was also confirmed by quantitative real-time PCR. Our data provide important clues for identifying and validating trans-activators that activate the gamma-globin gene in fetuses, and trans-acting factors involved in silencing the gamma-globin gene in adults.
Ma
Yan-Ni
YN
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, 5 Dong Dan San Tiao, Beijing 100005, People's Republic of China.
Zhang
Xin
X
Zhang
Jun-Wu
JW
Zhang
Xin-Hua
XH
Wang
Rong-Xin
RX
D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D004920:Erythropoiesis / Q000235:genetics; D020224:Expressed Sequence Tags; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D005914:Globins / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D007231:Infant, Newborn; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000378:metabolism*
D018952:Antigens, CD34; D017931:DNA Primers; D012333:RNA, Messenger; D015534:Trans-Activators; D005914:Globins