TY - JOUR
T1 - Identification of a genomic DNA sequence that quantitatively modulates KLF1 transcription factor expression in differentiating human hematopoietic cells
AU - Gnanapragasam, M. N.
AU - Planutis, A.
AU - Glassberg, J. A.
AU - Bieker, J. J.
N1 - Funding Information:
We thank Dr. Mignon Loh (UCSF) for JMML patient DNA samples. We thank Drs Barbara Graves and Kathleen Clark (University of Utah) for antibodies and discussion related to ETV6, and Dr. John Pimanda (Prince of Wales Hospital) for target binding sequence information. We thank Drs. Dan Bauer and Divya Vinjamur (Bauer lab; Harvard University), Scott Peslak and Aoi Wakabayashi (Blobel lab; University of Pennsylvania), and Laura Breda (Rivella lab; University of Pennsylvania) for cell culture protocol advice. We acknowledge Dr. Kaustav Mukherjee (MSSM) for ongoing discussions. We thank Drs. Vesna Najfeld and Joseph Tripodi (MSSM) for karyotype analysis, and Drs. Vivian Simon and Lubbertus Mulder (MSSM) for Amaxa usage. This work was supported by Doris Duke Charitable Foundation award 2019113 to JJB and JAG, by PHS grant R01 DK46865 to JJB, and by a Cooley’s Anemia Foundation Fellowship to MNG. The Yale Cooperative Center of Excellence in Hematology is supported by DK106829.
Funding Information:
We thank Dr. Mignon Loh (UCSF) for JMML patient DNA samples. We thank Drs Barbara Graves and Kathleen Clark (University of Utah) for antibodies and discussion related to ETV6, and Dr. John Pimanda (Prince of Wales Hospital) for target binding sequence information. We thank Drs. Dan Bauer and Divya Vinjamur (Bauer lab; Harvard University), Scott Peslak and Aoi Wakabayashi (Blobel lab; University of Pennsylvania), and Laura Breda (Rivella lab; University of Pennsylvania) for cell culture protocol advice. We acknowledge Dr. Kaustav Mukherjee (MSSM) for ongoing discussions. We thank Drs. Vesna Najfeld and Joseph Tripodi (MSSM) for karyotype analysis, and Drs. Vivian Simon and Lubbertus Mulder (MSSM) for Amaxa usage. This work was supported by Doris Duke Charitable Foundation award 2019113 to JJB and JAG, by PHS grant R01 DK46865 to JJB, and by a Cooley’s Anemia Foundation Fellowship to MNG. The Yale Cooperative Center of Excellence in Hematology is supported by DK106829.
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/12
Y1 - 2023/12
N2 - The onset of erythropoiesis is under strict developmental control, with direct and indirect inputs influencing its derivation from the hematopoietic stem cell. A major regulator of this transition is KLF1/EKLF, a zinc finger transcription factor that plays a global role in all aspects of erythropoiesis. Here, we have identified a short, conserved enhancer element in KLF1 intron 1 that is important for establishing optimal levels of KLF1 in mouse and human cells. Chromatin accessibility of this site exhibits cell-type specificity and is under developmental control during the differentiation of human CD34+ cells towards the erythroid lineage. This site binds GATA1, SMAD1, TAL1, and ETV6. In vivo editing of this region in cell lines and primary cells reduces KLF1 expression quantitatively. However, we find that, similar to observations seen in pedigrees of families with KLF1 mutations, downstream effects are variable, suggesting that the global architecture of the site is buffered towards keeping the KLF1 genetic region in an active state. We propose that modification of intron 1 in both alleles is not equivalent to complete loss of function of one allele.
AB - The onset of erythropoiesis is under strict developmental control, with direct and indirect inputs influencing its derivation from the hematopoietic stem cell. A major regulator of this transition is KLF1/EKLF, a zinc finger transcription factor that plays a global role in all aspects of erythropoiesis. Here, we have identified a short, conserved enhancer element in KLF1 intron 1 that is important for establishing optimal levels of KLF1 in mouse and human cells. Chromatin accessibility of this site exhibits cell-type specificity and is under developmental control during the differentiation of human CD34+ cells towards the erythroid lineage. This site binds GATA1, SMAD1, TAL1, and ETV6. In vivo editing of this region in cell lines and primary cells reduces KLF1 expression quantitatively. However, we find that, similar to observations seen in pedigrees of families with KLF1 mutations, downstream effects are variable, suggesting that the global architecture of the site is buffered towards keeping the KLF1 genetic region in an active state. We propose that modification of intron 1 in both alleles is not equivalent to complete loss of function of one allele.
UR - http://www.scopus.com/inward/record.url?scp=85158855472&partnerID=8YFLogxK
U2 - 10.1038/s41598-023-34805-5
DO - 10.1038/s41598-023-34805-5
M3 - Article
C2 - 37165057
AN - SCOPUS:85158855472
SN - 2045-2322
VL - 13
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 7589
ER -