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Erschienen in: Journal of Assisted Reproduction and Genetics 5/2016

06.04.2016 | Technological Innovations

Introducing precise genetic modifications into human 3PN embryos by CRISPR/Cas-mediated genome editing

verfasst von: Xiangjin Kang, Wenyin He, Yuling Huang, Qian Yu, Yaoyong Chen, Xingcheng Gao, Xiaofang Sun, Yong Fan

Erschienen in: Journal of Assisted Reproduction and Genetics | Ausgabe 5/2016

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Abstract

Purpose

As a powerful technology for genome engineering, the CRISPR/Cas system has been successfully applied to modify the genomes of various species. The purpose of this study was to evaluate the technology and establish principles for the introduction of precise genetic modifications in early human embryos.

Methods

3PN zygotes were injected with Cas9 messenger RNA (mRNA) (100 ng/μl) and guide RNA (gRNA) (50 ng/μl). For oligo-injections, donor oligo-1 (99 bp) or oligo-2 (99 bp) (100 ng/μl) or dsDonor (1 kb) was mixed with Cas9 mRNA (100 ng/μl) and gRNA (50 ng/μl) and injected into the embryos.

Results

By co-injecting Cas9 mRNA, gRNAs, and donor DNA, we successfully introduced the naturally occurring CCR5Δ32 allele into early human 3PN embryos. In the embryos containing the engineered CCR5Δ32 allele, however, the other alleles at the same locus could not be fully controlled because they either remained wild type or contained indel mutations.

Conclusions

This work has implications for the development of therapeutic treatments of genetic disorders, and it demonstrates that significant technical issues remain to be addressed. We advocate preventing any application of genome editing on the human germline until after a rigorous and thorough evaluation and discussion are undertaken by the global research and ethics communities.
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Metadaten
Titel
Introducing precise genetic modifications into human 3PN embryos by CRISPR/Cas-mediated genome editing
verfasst von
Xiangjin Kang
Wenyin He
Yuling Huang
Qian Yu
Yaoyong Chen
Xingcheng Gao
Xiaofang Sun
Yong Fan
Publikationsdatum
06.04.2016
Verlag
Springer US
Erschienen in
Journal of Assisted Reproduction and Genetics / Ausgabe 5/2016
Print ISSN: 1058-0468
Elektronische ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-016-0710-8

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