Subjects: Biology >> Biophysics >> Cell Biology submitted time 2016-05-12
Wang, Lixia
Liu, Guang-Hui
Wu, Jun
Belmonte, Juan Carlos Izpisua
Fang, Weiwei
Liu, Guang-Hui
Liu, Guang-Hui
Abstract： The CRISPR/Cas system has proven to be a powerful gene editing tool both in vitro and in vivo. A recent flurry of studies of in vivo gene editing using the CRISPR/Cas system bring bright prospects in creating animal models and targeted gene therapy of human genetic diseases.
Peer Review Status:
Awaiting Review
Subjects: Biology >> Biophysics >> Cell Biology submitted time 2016-05-12
Deng, Liping
Ren, Ruotong
Liu, Guang-Hui
Wu, Jun
Suzuki, Keiichiro
Belmote, Juan Carlos Izpisua
Liu, Guang-Hui
Liu, Guang-Hui
Abstract： Nuclease-based genome editing has proven to be a powerful and promising tool for disease modeling and gene therapy. Recent advances in CRISPR/Cas and TALE indicate that they could also be used as a targeted regulator of gene expression, as well as being utilized for illuminating specific chromosomal structures or genomic regions.
Peer Review Status:Awaiting Review
Subjects: Biology >> Biophysics >> Cell Biology submitted time 2016-05-12
Xie, Qing
Li, Xiangrui
Xie, Qing
Parlet, Corey
Borcherding, Nicholas
Kolb, Ryan
Li, Wei
Tygrett, Lorraine
Waldschmidt, Thomas
Olivier, Alicia
Zhang, Weizhou
Klesney-Tait, Julia
Keck, Kathy
Chen, Songhai
Liu, Guang-Hui
Liu, Guang-Hui
Abstract： Neutrophils play an essential role in the innate immune response to infection. Neutrophils migrate from the vasculature into the tissue in response to infection. Recently, a neutrophil cell surface receptor, CD177, was shown to help mediate neutrophil migration across the endothelium through interactions with PECAM1. We examined a publicly available gene array dataset of CD177 expression from human neutrophils following pulmonary endotoxin instillation. Among all 22,214 genes examined, CD177 mRNA was the most upregulated following endotoxin exposure. The high level of CD177 expression is also maintained in airspace neutrophils, suggesting a potential involvement of CD177 in neutrophil infiltration under infectious diseases. To determine the role of CD177 in neutrophils in vivo, we constructed a CD177-genetic knockout mouse model. The mice with homozygous deletion of CD177 have no discernible phenotype and no significant change in immune cells, other than decreased neutrophil counts in peripheral blood. We examined the role of CD177 in neutrophil accumulation using a skin infection model with Staphylococcus aureus. CD177 deletion reduced neutrophil counts in inflammatory skin caused by S. aureus. Mechanistically we found that CD177 deletion in mouse neutrophils has no significant impact in CXCL1/KC- or fMLP-induced migration, but led to significant cell death. Herein we established a novel genetic mouse model to study the role of CD177 and found that CD177 plays an important role in neutrophils.
Peer Review Status:Awaiting Review
Subjects: Biology >> Biophysics >> Biochemistry & Molecular Biology submitted time 2016-05-12
Reddy, Pradeep
Ocampo, Alejandro
Suzuki, Keiichiro
Luo, Jinping
Sugawara, Atsushi
Okamura, Daiji
Wu, Jun
Lam, David
Esteban, Concepcion Rodriguez
Sancho-Martinez, Ignacio
Belmonte, Juan Carlos Izpisua
Bacman, Sandra R.
Williams, Sion L.
Moraes, Carlos T.
Tsunekawa, Yuji
Xiong, Xiong
Zhao, Huimin
Montserrat, Nuria
Liu, Guang-Hui
Liu, Guang-Hui
Abstract： Mitochondrial diseases include a group of maternally inherited genetic disorders caused by mutations in mtDNA. In most of these patients, mutated mtDNA coexists with wild-type mtDNA, a situation known as mtDNA heteroplasmy. Here, we report on a strategy toward preventing germline transmission of mitochondrial diseases by inducing mtDNA heteroplasmy shift through the selective elimination of mutated mtDNA. As a proof of concept, we took advantage of NZB/BALB heteroplasmic mice, which contain two mtDNA haplotypes, BALB and NZB, and selectively prevented their germline transmission using either mitochondria-targeted restriction endonucleases or TALENs. In addition, we successfully reduced human mutated mtDNA levels responsible for Leber's hereditary optic neuropathy (LHOND), and neurogenic muscle weakness, ataxia, and retinitis pigmentosa (NARP), in mammalian oocytes using mitochondria-targeted TALEN (mito-TALENs). Our approaches represent a potential therapeutic avenue for preventing the trans-generational transmission of human mitochondrial diseases caused by mutations in mtDNA.
Peer Review Status:Awaiting Review
Subjects: Biology >> Biophysics >> Cell Biology submitted time 2016-05-05
Fu, Lina
Xu, Xiuling
Ren, Ruotong
Zhang, Weiqi
Yang, Jiping
Ren, Xiaoqing
Wang, Si
Zhao, Yang
Liu, Guang-Hui
Ren, Ruotong
Zhang, Weiqi
Liu, Guang-Hui
Qu, Jing
Wu, Jun
Belmonte, Juan Carlos Izpisua
Wu, Jun
Sun, Liang
Yang, Ze
Yu, Yang
Qiao, Jie
Abstract： Xeroderma pigmentosum (XP) is a group of genetic disorders caused by mutations of XP-associated genes, resulting in impairment of DNA repair. XP patients frequently exhibit neurological degeneration, but the underlying mechanism is unknown, in part due to lack of proper disease models. Here, we generated patient-specific induced pluripotent stem cells (iPSCs) harboring mutations in five different XP genes including XPA, XPB, XPC, XPG, and XPV. These iPSCs were further differentiated to neural cells, and their susceptibility to DNA damage stress was investigated. Mutation of XPA in either neural stem cells (NSCs) or neurons resulted in severe DNA damage repair defects, and these neural cells with mutant XPA were hyper-sensitive to DNA damage-induced apoptosis. Thus, XP-mutant neural cells represent valuable tools to clarify the molecular mechanisms of neurological abnormalities in the XP patients.
Peer Review Status:Awaiting Review
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