Arrayed functional genetic screenings in pluripotency reprogramming and differentiation

Author(s)

Panepucci, Rodrigo Alexandre ^{[1, 2]} ; de Souza Lima, Ildercilio Mota ^{[1, 2]}

Affiliation

^[1] Reg Blood Ctr Ribeirao Preto, Ctr Cell Based Therapy CTC, Lab Funct Biol LFBio, Rua Tenente Catao Roxo 2501, BR-14051140 Ribeirao Preto, SP - Brazil

^[2] Univ Sao Paulo, FMRP, Ribeirao Preto Med Sch, Dept Genet, Ribeirao Preto, SP - Brazil

• Document type: Review article
• Source: STEM CELL RESEARCH & THERAPY; v. 10, JAN 11 2019.
• Web of Science Citations: 1

Abstract

Thoroughly understanding the molecular mechanisms responsible for the biological properties of pluripotent stem cells, as well as for the processes involved in reprograming, differentiation, and transition between Naive and Primed pluripotent states, is of great interest in basic and applied research. Although pluripotent cells have been extensively characterized in terms of their transcriptome and miRNome, a comprehensive understanding of how these gene products specifically impact their biology, depends on gain- or loss-of-function experimental approaches capable to systematically interrogate their function. We review all studies carried up to date that used arrayed screening approaches to explore the function of these genetic elements on those biological contexts, using focused or genome-wide genetic libraries. We further discuss the limitations and advantages of approaches based on assays with population-level primary readouts, derived from single-parameter plate readers, or cell-level primary readouts, obtained using multiparametric flow cytometry or quantitative fluorescence microscopy (i.e., high-content screening). Finally, we discuss technical limitation and future perspectives, highlighting how the integration of screening data may lead to major advances in the field of stem cell research and therapy. (AU)

• FAPESP's process: 15/15864-6 - Functional evaluation of microRNAs in cell proliferation and differentiation of embryonal carcinoma cell line NTera-2 using high content screening
• Grantee: Ildercílio Mota de Souza Lima
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 15/08070-3 - Large-scale analysis of microRNA function in cell cycle, pluripotency, self-renewal and differentiation of stem cells using high-content screening
• Grantee: Rodrigo Alexandre Panepucci
• Support Opportunities: Scholarships abroad - Research
• FAPESP's process: 13/27061-0 - The role of microRNAs on the proliferation and cell differentiation of the embryonal carcinoma pluripotent cell line NTera-2.
• Grantee: Ildercílio Mota de Souza Lima
• Support Opportunities: Scholarships in Brazil - Doctorate