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Gene correction. Human 2PN model
• CRISPR/Cas9 is effective as a gene-editing tool in human 2PN zygotes.
• By injection of Cas9 protein complexed with the appropriate sgRNAs and homology donors into one- cell human embryos, efficient homologous recombination-mediated correction of point mutations in HBB and G6PD was demonstrated.
CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein Mol Genet. Genomics, Tang, 2017
PCC Genetics - ESHRE BCN 2018
Gene correction. Human 2 PN model
• Correction of the heterozygous MYBPC3 mutation in human preimplantation embryos with precise CRISPR–Cas9-based targeting accuracy and high homology-directed repair efficiency.
• Endogenous, germline-specific DNA repair response.
• Induced double-strand breaks (DSBs) at the mutant paternal allele were predominantly repaired using the homologous wild-type maternal gene instead of a synthetic DNA template.
• By modulating the cell cycle stage at which the DSB was induced, mosaicism was avoided in cleaving embryos.
Ruzo and Bivanlou, 2017
Correction of a pathogenic gene mutation in human embryos Nature, Ma et al, 2017 PCC Genetics - ESHRE BCN 2018
Injection of Cas9 protein and sgRNA together with the sperm, at the time ICSI. Correction of 45% of the mutant embryos completely, with no mosaicism and no detectable off-targets.
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