Correction of a pathogenic gene mutation in human embryos

Published on Aug 1, 2017in Nature43.07
· DOI :10.1038/nature23305
Hong Ma27
Estimated H-index: 27
(Oregon National Primate Research Center),
Nuria Marti-Gutierrez1
Estimated H-index: 1
(OHSU: Oregon Health & Science University)
+ 28 AuthorsShoukhrat Mitalipov36
Estimated H-index: 36
CRISPR–Cas9 genome editing is used to induce a DNA repair response and correct a disease-causing heterozygous mutation in human embryos with reduced mosaicism and preferential repair using the wild-type copy of the gene.
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  • References (48)
  • Citations (259)
Published on Jun 1, 2017in Molecular Genetics and Genomics2.88
Lichun Tang1
Estimated H-index: 1
(Protein Sciences),
Yanting Zeng1
Estimated H-index: 1
(Guangzhou Medical University)
+ 8 AuthorsXiaowei Li1
Estimated H-index: 1
(NU: Nanjing University)
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 o...
Published on Feb 1, 2017in Scientific Reports4.01
Zhuchi Tu7
Estimated H-index: 7
Weili Yang9
Estimated H-index: 9
+ 10 AuthorsSu Yang9
Estimated H-index: 9
Published on Jan 15, 2017in Bioinformatics4.53
Jeongbin Park8
Estimated H-index: 8
(Hanyang University),
Kayeong Lim4
Estimated H-index: 4
(SNU: Seoul National University)
+ 1 AuthorsSangsu Bae13
Estimated H-index: 13
(Hanyang University)
Published on Oct 1, 2016in Developmental Biology2.94
Masakazu Hashimoto7
Estimated H-index: 7
(Osaka University),
Yukiko Yamashita2
Estimated H-index: 2
(University of Tokushima),
Tatsuya Takemoto13
Estimated H-index: 13
(University of Tokushima)
Abstract The CRISPR/Cas9 system is a powerful tool for elucidating the roles of genes in a wide variety of organisms including mice. To obtain genetically modified embryos or mice by this method, Cas9 mRNA and sgRNA are usually introduced into zygotes by microinjection or electroporation. However, most mutants generated with this method are genetically mosaic, composed of several types of cells carrying different mutations, which complicates phenotype analysis in founder embryos or mice. To simp...
Published on Oct 1, 2016in Cell36.22
Julian Lange12
Estimated H-index: 12
(MSK: Memorial Sloan Kettering Cancer Center),
Shintaro Yamada3
Estimated H-index: 3
(MSK: Memorial Sloan Kettering Cancer Center)
+ 6 AuthorsScott Keeney47
Estimated H-index: 47
(Cornell University)
Summary Heritability and genome stability are shaped by meiotic recombination, which is initiated via hundreds of DNA double-strand breaks (DSBs). The distribution of DSBs throughout the genome is not random, but mechanisms molding this landscape remain poorly understood. Here, we exploit genome-wide maps of mouse DSBs at unprecedented nucleotide resolution to uncover previously invisible spatial features of recombination. At fine scale, we reveal a stereotyped hotspot structure—DSBs occur withi...
Published on May 1, 2016in Cell Stem Cell21.46
Eunju Kang13
Estimated H-index: 13
(Oregon National Primate Research Center),
Xinjian Wang20
Estimated H-index: 20
(Cincinnati Children's Hospital Medical Center)
+ 20 AuthorsYing Li14
Estimated H-index: 14
(Oregon National Primate Research Center)
Summary The genetic integrity of iPSCs is an important consideration for therapeutic application. In this study, we examine the accumulation of somatic mitochondrial genome (mtDNA) mutations in skin fibroblasts, blood, and iPSCs derived from young and elderly subjects (24–72 years). We found that pooled skin and blood mtDNA contained low heteroplasmic point mutations, but a panel of ten individual iPSC lines from each tissue or clonally expanded fibroblasts carried an elevated load of heteroplas...
Published on Mar 1, 2016in Nature Biotechnology31.86
Christopher D. Richardson27
Estimated H-index: 27
Graham Jordan Ray4
Estimated H-index: 4
+ 2 AuthorsJacob E. Corn29
Estimated H-index: 29
The efficiency of homology-directed genome editing with CRISPR-Cas9 is boosted through improved design of donor DNA.
Published on Mar 1, 2016in Genome Research9.94
Daesik Kim12
Estimated H-index: 12
(SNU: Seoul National University),
Sojung Kim10
Estimated H-index: 10
(SNU: Seoul National University)
+ 2 AuthorsJin-Soo Kim52
Estimated H-index: 52
(SNU: Seoul National University)
We present multiplex Digenome-seq to profile genome-wide specificities of up to 11 CRISPR-Cas9 nucleases simultaneously, saving time and reducing cost. Cell-free human genomic DNA was digested using multiple sgRNAs combined with the Cas9 protein and then subjected to whole-genome sequencing. In vitro cleavage patterns, characteristic of on- and off-target sites, were computationally identified across the genome using a new DNA cleavage scoring system. We found that many false-positive, bulge-typ...
Published on Dec 20, 2015in Human Molecular Genetics4.54
Yu Kang8
Estimated H-index: 8
(Kunming University),
Bo Zheng8
Estimated H-index: 8
(Nanjing Medical University)
+ 13 AuthorsHong Wang9
Estimated H-index: 9
(Kunming University)
Mutations in the DAX1 locus cause X-linked adrenal hypoplasia congenita (AHC) and hypogonadotropic hypogonadism (HH), which manifest with primary adrenal insufficiency and incomplete or absent sexual maturation, respectively. The associated defects in spermatogenesis can range from spermatogenic arrest to Sertoli cell only syndrome. Conclusions from Dax1 knockout mouse models provide only limited insight into AHC/HH disease mechanisms, because mouse models exhibit more extensive abnormalities in...
Published on Dec 1, 2015in Gene2.64
Lucie Carrier43
Estimated H-index: 43
(UHH: University of Hamburg),
Giulia Mearini16
Estimated H-index: 16
(UHH: University of Hamburg)
+ 1 AuthorsFriederike Cuello6
Estimated H-index: 6
(UHH: University of Hamburg)
Abstract More than 350 individual MYPBC3 mutations have been identified in patients with inherited hypertrophic cardiomyopathy (HCM), thus representing 40–50% of all HCM mutations, making it the most frequently mutated gene in HCM. HCM is considered a disease of the sarcomere and is characterized by left ventricular hypertrophy, myocyte disarray and diastolic dysfunction. MYBPC3 encodes for the thick filament associated protein cardiac myosin-binding protein C (cMyBP-C), a signaling node in card...
Cited By259
Published on Dec 1, 2019in BMC Biotechnology2.30
Kayla Carey (VT: Virginia Tech), Junghyun Ryu5
Estimated H-index: 5
(VT: Virginia Tech)
+ 4 AuthorsKiho Lee6
Estimated H-index: 6
(VT: Virginia Tech)
Background The CRISPR/Cas9 system can effectively introduce site-specific modifications to the genome. The efficiency is high enough to induce targeted genome modifications during embryogenesis, thus increasing the efficiency of producing genetically modified animal models and having potential clinical applications as an assisted reproductive technology. Because most of the CRISPR/Cas9 systems introduce site-specific double-stranded breaks (DSBs) to induce site-specific modifications, a major co...
Published on Jan 8, 2019in Nature Communications11.88
Puping Liang7
Estimated H-index: 7
(SYSU: Sun Yat-sen University),
Xiaowei Xie6
Estimated H-index: 6
(SYSU: Sun Yat-sen University)
+ 9 AuthorsDan Liu26
Estimated H-index: 26
(BCM: Baylor College of Medicine)
The adenine base editor (ABE), capable of catalyzing A•T to G•C conversions, is an important gene editing toolbox. Here, we systematically evaluate genome-wide off-target deamination by ABEs using the EndoV-seq platform we developed. EndoV-seq utilizes Endonuclease V to nick the inosine-containing DNA strand of genomic DNA deaminated by ABE in vitro. The treated DNA is then whole-genome sequenced to identify off-target sites. Of the eight gRNAs we tested with ABE, 2–19 (with an average of 8.0) o...
Published on Jul 16, 2018in Insect Science2.71
Sanyuan Ma13
Estimated H-index: 13
(SWU: Southwest University),
Guy Smagghe51
Estimated H-index: 51
(UGent: Ghent University),
Estimated H-index: 38
(SWU: Southwest University)
Published on Jul 1, 2019in Neurobiology of Disease5.16
Udaiyappan Janakiraman (UA: University of Arizona), Jie Yu1
Estimated H-index: 1
(ZJNU: Zhejiang Chinese Medical University)
+ 6 AuthorsMark A. Nelson32
Estimated H-index: 32
(UA: University of Arizona)
Abstract TAF1/MRSX33 intellectual disability syndrome is an X-linked disorder caused by loss-of-function mutations in the TAF1 gene. How these mutations cause dysmorphology, hypotonia, intellectual and motor defects is unknown. Mouse models which have embryonically targeted TAF1 have failed, possibly due to TAF1 being essential for viability, preferentially expressed in early brain development, and intolerant of mutation. Novel animal models are valuable tools for understanding neuronal patholog...
Published on Dec 1, 2019in Genome Biology14.03
Meiling Zhang2
Estimated H-index: 2
(SJTU: Shanghai Jiao Tong University),
Changyang Zhou3
Estimated H-index: 3
(CAS: Chinese Academy of Sciences)
+ 14 AuthorsNing Yao2
Estimated H-index: 2
(SJTU: Shanghai Jiao Tong University)
Base editing installs a precise nucleotide change in specific gene loci without causing a double-strand break. Its efficiency in human embryos is generally low, limiting its utility in functional genetic studies. Here, we report that injecting base editors into human cleaving two-cell and four-cell embryos results in much higher (up to 13-fold) homozygotic nucleotide substitution efficiency as opposed to MII oocytes or zygotes. Furthermore, as a proof-of-principle study, a point mutation can be ...
Published on Jun 1, 2019in Molecular therapy. Nucleic acids5.92
Guanglei Li4
Estimated H-index: 4
(Guangzhou Medical University),
Xinyi Liu (JNU: Jinan University)+ 8 AuthorsXingxu Huang14
Estimated H-index: 14
(CAS: Chinese Academy of Sciences)
Base editing systems show their power in modeling and correcting the pathogenic mutations of genetic diseases. Previous studies have already demonstrated the editing efficiency of BE3-mediated C-to-T conversion in human embryos. However, the precision and efficiency of a recently developed adenine base editor (ABE), which converts A-to-G editing in human embryos, remain to be addressed. Here we selected reported pathogenic mutations to characterize the ABE in human tripronuclear embryos. We foun...
Erika Kleiderman5
Estimated H-index: 5
(McGill University),
Ian Stedman (York University)
Human germline genome editing may prove to be especially poignant for members of the rare disease community, many of whom are diagnosed with monogenic diseases. This community lacks broad representation in the literature surrounding genome editing, notably in Canada, yet is likely to be directly affected by eventual clinical applications of this technology. Although not generalizable, the literature does offer some commonalities regarding the experiences of rare disease patients. This manuscript...
Published in bioRxiv
Koray Dogan Kaya6
Estimated H-index: 6
(NIH: National Institutes of Health),
Hsiu-Chuan Chen2
Estimated H-index: 2
(NIH: National Institutes of Health)
+ -3 AuthorsK. Kruczek (NIH: National Institutes of Health)
Retinal organoids generated from human pluripotent stem cells exhibit considerable variability in temporal dynamics of differentiation. To assess the maturity of neural retina in vitro, we performed transcriptome analyses of developing organoids from human embryonic and induced pluripotent stem cell lines. We show that the developmental variability in organoids was reflected in gene expression profiles and could be evaluated by molecular staging with the human fetal and adult retinal transcripto...
Published on May 1, 2019in European Journal of Medical Genetics2.02
Joyce C. Harper46
Estimated H-index: 46
(UCL: University College London),
Gerald Schatten71
Estimated H-index: 71
(University of Pittsburgh)
Abstract Perhaps the two most significant pioneering biomedical discoveries with immediate clinical implications during the past forty years have been the advent of assisted reproductive technologies (ART) and the genetics revolution. ART, including in vitro fertilization (IVF), intracytoplasmic sperm injection and preimplantation genetic testing, has resulted in the birth of more than 8 million children, and the pioneer of IVF, Professor Bob Edwards, was awarded the 2010 Nobel Prize. The geneti...
Published in DNA Repair3.71
Sandra Bosshard2
Estimated H-index: 2
(UNIL: University of Lausanne),
Pierre-Olivier Duroy (UNIL: University of Lausanne), Nicolas Mermod30
Estimated H-index: 30
(UNIL: University of Lausanne)
Abstract CRISPR technologies greatly foster genome editing in mammalian cells through site-directed DNA double strand breaks (DSBs). However, precise editing outcomes, as mediated by homologous recombination (HR) repair, are typically infrequent and outnumbered by undesired genome alterations. By using knockdown and overexpression studies in Chinese hamster ovary (CHO) cells as well as characterizing repaired DNA junctions, we found that efficient HR-mediated genome editing depends on alternativ...