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

Author(s)

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

Affiliation

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

• Document type: Journal article
• Source: STEM CELL RESEARCH; v. 31, p. 52-54, AUG 2018.
• Web of Science Citations: 1

Abstract

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)

• FAPESP's process: 16/22318-0 - Alternative approaches targeting Duchenne Muscular Dystrophy treatment
• Grantee: Felipe de Souza Leite
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 16/14517-3 - Search for "protective" modifying variants/mechanisms in asymptomatic or very mildly affected individuals with pathogenic mutation in SPAST gene
• Grantee: Uirá Souto Melo
• Support type: Scholarships in Brazil - Post-Doctorate
• FAPESP's process: 13/08028-1 - CEGH-CEL - Human Genome and Stem Cell Research Center
• Grantee: Mayana Zatz
• Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC