High-Throughput siRNA Screening to Reveal GATA-2 Upstream Transcriptional Mechanisms in Hematopoietic Cells

PloS One
Yo SaitoHideo Harigae

Abstract

Hematopoietic stem cells can self-renew and differentiate into all blood cell types. The transcription factor GATA-2 is expressed in both hematopoietic stem and progenitor cells and is essential for cell proliferation, survival, and differentiation. Recently, evidence from studies of aplastic anemia, MonoMAC syndrome, and lung cancer has demonstrated a mechanistic link between GATA-2 and human pathophysiology. GATA-2-dependent disease processes have been extensively analyzed; however, the transcriptional mechanisms upstream of GATA-2 remain less understood. Here, we conducted high-throughput small-interfering-RNA (siRNA) library screening and showed that YN-1, a human erythroleukemia cell line, expressed high levels of GATA-2 following the activation of the hematopoietic-specific 1S promoter. As transient luciferase reporter assay in YN-1 cells revealed the highest promoter activity in the 1S promoter fused with GATA-2 intronic enhancer (+9.9 kb/1S); therefore, we established a cell line capable of stably expressing +9.9 kb/1S-Luciferase. Subsequently, we screened 995 transcription factor genes and revealed that CITED2 acts as a GATA-2 activator in human hematopoietic cells. These results provide novel insights into and further...Continue Reading

References

Aug 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·T EvansG Felsenfeld
Jan 1, 1980·Leukemia Research·H P KoefflerD W Golde
Mar 7, 1998·The Journal of Biological Chemistry·N MinegishiM Yamamoto
Apr 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·G A BlobelS H Orkin
Oct 16, 2003·Molecular and Cellular Biology·Kamil R KrancShoumo Bhattacharya
Sep 9, 2006·The Tohoku Journal of Experimental Medicine·Hideo Harigae
Mar 31, 2007·Development·Melin KhandekarJames Douglas Engel
Feb 26, 2008·Cell·Stuart H Orkin, Leonard I Zon
Dec 3, 2009·Cell Stem Cell·Kamil R KrancTariq Enver
Jul 31, 2010·The Journal of Biological Chemistry·Emery H BresnickSunduz Keles
Sep 15, 2010·PLoS Genetics·Jonathan W SnowEmery H Bresnick
Mar 15, 2011·Blood·Jonathan W SnowEmery H Bresnick
May 7, 2011·Stem Cells and Development·Xiaolei DingMartin Zenke
Sep 22, 2012·The Journal of Clinical Investigation·Kirby D JohnsonEmery H Bresnick
Dec 26, 2012·The Journal of Experimental Medicine·Jordi GuiuAnna Bigas
Mar 13, 2014·Proceedings of the National Academy of Sciences of the United States of America·Rajendran SanalkumarEmery H Bresnick
Mar 19, 2014·International Journal of Hematology·Takashi Moriguchi, Masayuki Yamamoto
Aug 26, 2014·Haematologica·Mayumi KamataHideo Harigae

❮ Previous
Next ❯

Citations

Jun 2, 2017·The Tohoku Journal of Experimental Medicine·Tohru Fujiwara
Feb 22, 2017·Biochemical and Biophysical Research Communications·Tohru FujiwaraHideo Harigae
Jan 19, 2018·The Tohoku Journal of Experimental Medicine·Keiichi OhashiHideo Harigae

❮ Previous
Next ❯

Methods Mentioned

BETA
Assay
transfection
electrophoresis

Related Concepts

Related Feeds

Aminoglycosides (ASM)

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is a clinically and genetically heterogeneous disease with approximately 20,000 cases per year in the United States. AML also accounts for 15-20% of all childhood acute leukemias, while it is responsible for more than half of the leukemic deaths in these patients. Here is the latest research on this disease.

CREs: Gene & Cell Therapy

Gene and cell therapy advances have shown promising outcomes for several diseases. The role of cis-regulatory elements (CREs) is crucial in the design of gene therapy vectors. Here is the latest research on CREs in gene and cell therapy.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

Blood And Marrow Transplantation

The use of hematopoietic stem cell transplantation or blood and marrow transplantation (bmt) is on the increase worldwide. BMT is used to replace damaged or destroyed bone marrow with healthy bone marrow stem cells. Here is the latest research on bone and marrow transplantation.

Adult Stem Cells

Adult stem cells reside in unique niches that provide vital cues for their survival, self-renewal, and differentiation. They hold great promise for use in tissue repair and regeneration as a novel therapeutic strategies. Here is the latest research.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

Birth Defects

Birth defects encompass structural and functional alterations that occur during embryonic or fetal development and are present since birth. The cause may be genetic, environmental or unknown and can result in physical and/or mental impairment. Here is the latest research on birth defects.

Aminoglycosides

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

AML: Role of LSD1 by CRISPR (Keystone)

Find the latest rersearrch on the ability of CRISPR-Cas9 mutagenesis to profile the interactions between lysine-specific histone demethylase 1 (LSD1) and chemical inhibitors in the context of acute myeloid leukemia (AML) here.

Acute Myeloid Leukaemia & RNA

Acute myeloid leukaemia (AML) is a common hematological type of cancer. As the population ages, there has been a rise in the frequency of AML. RNA expression has been used to see if there are different genetic profiles that exist within AML and whether these may underpin the variations in survival rates. Here is the latest research on AML and RNA.

Related Papers

[Rinshō ketsueki] The Japanese journal of clinical hematology
K Miura
[Rinshō ketsueki] The Japanese journal of clinical hematology
A Kanamaru, Y Hara
Proceedings of the National Academy of Sciences of the United States of America
L I ZonS H Orkin
© 2021 Meta ULC. All rights reserved