서브메뉴
검색
Dissecting Differences in Function of CBX Paralogs During Differentiation to Neural Progenitor Cells
Dissecting Differences in Function of CBX Paralogs During Differentiation to Neural Progenitor Cells
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211152800
- ISBN
- 9798346567226
- DDC
- 574
- 서명/저자
- Dissecting Differences in Function of CBX Paralogs During Differentiation to Neural Progenitor Cells
- 발행사항
- [Sl] : Harvard University, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 189 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 86-05, Section: B.
- 주기사항
- Advisor: Kingston, Robert E.
- 학위논문주기
- Thesis (Ph.D.)--Harvard University, 2024.
- 초록/해제
- 요약During development, a single cell gives rise to a complex multicellular organism through establishment and maintenance of cell type-specific gene expression. Polycomb-mediated silencing of lineage inappropriate genes is essential to this process. Knockout of canonical Polycomb Repressive Complex 1 (cPRC1) CBX family paralogs Cbx2 or Cbx4 results in postnatal lethality in mice. This is in contrast to other PcG proteins whose knockout causes early embryonic lethality. Notably, paralogs CBX2, CBX4, and CBX8 share similar domain structure and are co-expressed in differentiated cells. We hypothesize that CBX2, CBX4, and CBX8 are partially redundant while retaining unique functions.CBX2, CBX4, and CBX8 have shared biochemical activities, including nucleosome compaction and condensate formation. To investigate their shared functionality in cells, I utilized an in vitro differentiation system in which mouse embryonic stem cells (mESCs) are differentiated into neural progenitor cells (NPCs). I profiled expression of CBX2, CBX4, and CBX8 throughout differentiation. Expression at the transcript and protein levels shows distinct profiles for CBX2 versus CBX4 and CBX8, suggesting unique roles despite shared biochemical activities.I generated single knockout lines and found that knockout of a single CBX protein resulted in some shared transcriptional changes in NPCs, indicating overlapping function. However, CBX2 knockout caused unique transcriptional changes that signal an earlier arrest in differentiation. I then generated double knockouts, whose transcriptional phenotype was determined by presence or absence of CBX2. Therefore, loss of CBX2 is epistatic to loss of CBX4 or CBX8 during NPC differentiation. These double knockouts did not show a more severe transcriptional phenotype than relevant single knockout lines, so I generated triple knockout lines. Loss of all three paralogs had a morphological phenotype that was not rescued by exogenous expression of CBX2 or of CBX4 and CBX8 although partial transcriptional rescue was observed with exogenous expression of CBX2. Loss of all three paralogs might alter cell state by disrupting Polycomb function in differentiating lineages, providing further evidence of redundancy. In all, we demonstrate that paralogs CBX2 has a unique role in the exit from pluripotency while providing evidence for overlapping roles of CBX2, CBX4, and CBX8 in neural progenitor cell differentiation.
- 일반주제명
- Biology
- 일반주제명
- Cellular biology
- 일반주제명
- Molecular biology
- 키워드
- CBX paralogs
- 기타저자
- Harvard University Medical Sciences
- 기본자료저록
- Dissertations Abstracts International. 86-05B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
008250123s2024 us c eng d■001000017163840
■00520250211152800
■006m o d
■007cr#unu||||||||
■020 ▼a9798346567226
■035 ▼a(MiAaPQ)AAI31556377
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aMauger, MacKenzie.▼0(orcid)0000-0002-4536-8234
■24510▼aDissecting Differences in Function of CBX Paralogs During Differentiation to Neural Progenitor Cells
■260 ▼a[Sl]▼bHarvard University▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a189 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-05, Section: B.
■500 ▼aAdvisor: Kingston, Robert E.
■5021 ▼aThesis (Ph.D.)--Harvard University, 2024.
■520 ▼aDuring development, a single cell gives rise to a complex multicellular organism through establishment and maintenance of cell type-specific gene expression. Polycomb-mediated silencing of lineage inappropriate genes is essential to this process. Knockout of canonical Polycomb Repressive Complex 1 (cPRC1) CBX family paralogs Cbx2 or Cbx4 results in postnatal lethality in mice. This is in contrast to other PcG proteins whose knockout causes early embryonic lethality. Notably, paralogs CBX2, CBX4, and CBX8 share similar domain structure and are co-expressed in differentiated cells. We hypothesize that CBX2, CBX4, and CBX8 are partially redundant while retaining unique functions.CBX2, CBX4, and CBX8 have shared biochemical activities, including nucleosome compaction and condensate formation. To investigate their shared functionality in cells, I utilized an in vitro differentiation system in which mouse embryonic stem cells (mESCs) are differentiated into neural progenitor cells (NPCs). I profiled expression of CBX2, CBX4, and CBX8 throughout differentiation. Expression at the transcript and protein levels shows distinct profiles for CBX2 versus CBX4 and CBX8, suggesting unique roles despite shared biochemical activities.I generated single knockout lines and found that knockout of a single CBX protein resulted in some shared transcriptional changes in NPCs, indicating overlapping function. However, CBX2 knockout caused unique transcriptional changes that signal an earlier arrest in differentiation. I then generated double knockouts, whose transcriptional phenotype was determined by presence or absence of CBX2. Therefore, loss of CBX2 is epistatic to loss of CBX4 or CBX8 during NPC differentiation. These double knockouts did not show a more severe transcriptional phenotype than relevant single knockout lines, so I generated triple knockout lines. Loss of all three paralogs had a morphological phenotype that was not rescued by exogenous expression of CBX2 or of CBX4 and CBX8 although partial transcriptional rescue was observed with exogenous expression of CBX2. Loss of all three paralogs might alter cell state by disrupting Polycomb function in differentiating lineages, providing further evidence of redundancy. In all, we demonstrate that paralogs CBX2 has a unique role in the exit from pluripotency while providing evidence for overlapping roles of CBX2, CBX4, and CBX8 in neural progenitor cell differentiation.
■590 ▼aSchool code: 0084.
■650 4▼aBiology
■650 4▼aCellular biology
■650 4▼aMolecular biology
■653 ▼aCBX paralogs
■653 ▼aCanonical Polycomb Repressive Complex 1
■653 ▼aMouse embryonic stem cells
■690 ▼a0306
■690 ▼a0379
■690 ▼a0307
■71020▼aHarvard University▼bMedical Sciences.
■7730 ▼tDissertations Abstracts International▼g86-05B.
■790 ▼a0084
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17163840▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
