Arrayed functional genetic screenings in pluripotency reprogramming and differentiation

Autor(es)

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

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo de Revisão
• Fonte: STEM CELL RESEARCH & THERAPY; v. 10, JAN 11 2019.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 13/27061-0 - O papel dos microRNAs na proliferação e diferenciação celular da linhagem pluripotente de carcinoma embrionário humano NTera-2
• Beneficiário: Ildercílio Mota de Souza Lima
• Linha de fomento: Bolsas no Brasil - Doutorado
• Processo FAPESP: 15/08070-3 - Análise em larga-escala da função dos microRNAs no ciclo celular, pluripotência, auto-renovação e diferenciação de células-tronco utilizando "High Content Screening"
• Beneficiário: Rodrigo Alexandre Panepucci
• Linha de fomento: Bolsas no Exterior - Pesquisa
• Processo FAPESP: 15/15864-6 - Avaliação funcional de microRNAs na proliferação e diferenciação celular da linhagem de carcinoma embrionário NTera-2 por high content screening
• Beneficiário: Ildercílio Mota de Souza Lima
• Linha de fomento: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 13/08135-2 - CTC - Centro de Terapia Celular
• Beneficiário: Dimas Tadeu Covas
• Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs