CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein

@article{Tang2017CRISPRCas9mediatedGE,
  title={CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein},
  author={Lichun Tang and Yanting Zeng and Hongzi Du and Mengmeng Gong and Jin Peng and Buxi Zhang and Ming Lei and Fang Zhao and Weihua Wang and Xiaowei Li and Jian-qiao Liu},
  journal={Molecular Genetics and Genomics},
  year={2017},
  volume={292},
  pages={525-533}
}
Previous works using human tripronuclear zygotes suggested that the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system could be a tool in correcting disease-causing mutations. However, whether this system was applicable in normal human (dual pronuclear, 2PN) zygotes was unclear. Here we demonstrate that CRISPR/Cas9 is also effective as a gene-editing tool in human 2PN zygotes. By injection of Cas9 protein complexed with the appropriate sgRNAs and homology donors into… Expand
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References

SHOWING 1-10 OF 29 REFERENCES
CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes
TLDR
It is found that CRISPR/Cas9 could effectively cleave the endogenous β-globin gene (HBB), however, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic. Expand
Heritable Multiplex Genetic Engineering in Rats Using CRISPR/Cas9
TLDR
The CRISPR/Cas9 system makes it possible to efficiently and reliably generate gene knock-out rats by co-injection of Cas9 mRNA and sgRNAs into fertilized eggs and observed the gene modifications are germline transmittable. Expand
Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes*
TLDR
This work describes a simple and economic electroporation-based strategy to deliver Cas9/sgRNA ribonucleoproteins into mouse zygotes with 100% efficiency for in vivo genome editing and uses CRISPR-EZ to target the tyrosinase (Tyr) gene. Expand
Introducing precise genetic modifications into human 3PN embryos by CRISPR/Cas-mediated genome editing
TLDR
Evaluating the CRISPR/Cas technology and establishing principles for the introduction of precise genetic modifications in early human embryos demonstrates that significant technical issues remain to be addressed. Expand
Electroporation of Cas9 protein/sgRNA into early pronuclear zygotes generates non-mosaic mutants in the mouse.
TLDR
This method solves the problem of mosaicism/allele complexity in founder mutant embryos or mice generated by the CRIPSR/Cas9 system by introducing Cas9 protein and sgRNA into in vitro fertilized (IVF) zygotes by electroporation. Expand
One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering
TLDR
The CRISPR/Cas system allows the one-step generation of animals carrying mutations in multiple genes, an approach that will greatly accelerate the in vivo study of functionally redundant genes and of epistatic gene interactions. Expand
Multiplex Genome Engineering Using CRISPR/Cas Systems
TLDR
Two different type II CRISPR/Cas systems are engineered and it is demonstrated that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology. Expand
Generation of Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos
TLDR
By coinjection of Cas9 mRNA and sgRNAs into one-cell-stage embryos, this system successfully achieves precise gene targeting in cynomolgus monkeys and enables simultaneous disruption of two target genes in one step, and no off-target mutagenesis was detected by comprehensive analysis. Expand
RNA-Guided Human Genome Engineering via Cas9
TLDR
The type II bacterial CRISPR system is engineer to function with custom guide RNA (gRNA) in human cells to establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering. Expand
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