A fast method to reprogram and CRISPR/Cas9 gene editing from erythroblasts

Autor(es)

Melo, Uira Souto ^[1] ; Leite, Felipe de Souza ^[1] ; Costa, Silvia ^[1] ; Rosenberg, Carla ^[1] ; Zatz, Mayana ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Human Genome & Stem Cell Res Ctr, Biosci Inst, Dept Genet & Evolutionary Biol, BR-05508900 Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: STEM CELL RESEARCH; v. 31, p. 52-54, AUG 2018.
• Citações Web of Science: 1

Resumo

An efficient one-step procedure to reprogram fibroblasts into human induced pluripotent stem cells (hiPSC) and perform CRISPR/Cas9 gene editing simultaneously was recently reported. Here we show that such simultaneous reprogramming and gene editing can be efficiently done from erythroblasts. We successfully obtained human induced pluripotent stem cells colonies together with in frame and out of frame CAPN1 mutations in one or both alleles. We did not identify off-targets in edited cell lines. The entire process, from blood collection to mutated hiPSC took approximately 5 weeks, a much shorter period than standard multi-step methodologies using fibroblasts. Noteworthy, blood drawing is a less invasive procedure than a skin biopsy. (AU)

• Processo FAPESP: 16/22318-0 - Novas estratégias visando o tratamento da Distrofia de Duchenne
• Beneficiário: Felipe de Souza Leite
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/08028-1 - CEGH-CEL - Centro de Estudos do Genoma Humano e de Células-Tronco
• Beneficiário: Mayana Zatz
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 16/14517-3 - Identificação de variantes/mecanismos protetores em indivíduos com mutação patogênica no gene SPAST assintomáticos ou levemente afetados
• Beneficiário: Uirá Souto Melo
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado