ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding

Yantian District, China

ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding

Yantian District, China
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Meng Y.,Jinan University | Shi C.,Jinan University | Hu B.,Jinan University | Gong J.,Jinan University | And 6 more authors.
BMC Cell Biology | Year: 2017

Background: Mesenchymal stem cells (MSCs) are multipotent stromal cells that have the ability to self-renew and migrate to sites of pathology. In vivo tracking of MSCs provides insights into both, the underlying mechanisms of MSC transformation and their potential as gene delivery vehicles. The aim of our study was to assess the ability of superparamagnetic iron oxide nanoparticles (SPIONs)-labeled Wharton's Jelly of the human umbilical cord-derived MSCs (WJ-MSCs) to carry the green fluorescent protein (GFP) gene to cutaneous injury sites in a murine model. Methods: WJ-MSCs were isolated from a fresh umbilical cord and were genetically transformed to carry the GFP gene using lentiviral vectors with magnetically labeled SPIONs. The SPIONs/GFP-positive WJ-MSCs expressed multipotent cell markers and demonstrated the potential for osteogenic and adipogenic differentiation. Fifteen skininjured mice were divided into three groups. Group I was treated with WJ-MSCs, group II with SPIONs/GFP-positive WJ-MSCs, and group III with SPIONs/GFP-positive WJ-MSCs exposed to an external magnetic field (EMF). Magnetic resonance imaging and optical molecular imaging were performed, and images were acquired 1, 2, and 7 days after cell injection. Results: The results showed that GFP could be intensively detected around the wound in vivo 24 h after the cells were injected. Furthermore, we observed an accumulation of WJ-MSCs at the wound site, and EMF exposure increased the speed of cell transport. In conclusion, our study demonstrated that SPIONs/GFP function as cellular probes for monitoring in vivo migration and homing of WJ-MSCs. Moreover, exposure to an EMF can increase the transportation efficiency of SPIONs-labeled WJ-MSCs in vivo. Conclusions: Our findings could lead to the development of a gene carrier system for the treatment of diseases. © 2017 The Author(s).


PubMed | Shenzhen Engineering Laboratory for Genomics Assisted Animal Breeding, BGI ARK Biotechnology Co. and China; BGI ARK Biotechnology Co.
Type: Evaluation Studies | Journal: Yi chuan = Hereditas | Year: 2014

DNA editing techniques for targeted genome modification have witnessed remarkable advances and been widely used in various organisms. However, traditional gene targeting and cloning method has been shown to be low efficient, time-consuming and expensive for generating knockout animals, especially for big animals. Here we report the generation of site-specific genome modified pig with the newly developed artificially engineered sequence-specific endonucleases (transcription activator-like effector nuclease, TALENs) and handmade cloning (HMC) methods. First, we constructed the porcine GHR-knockout vector according to TALENs kit protocol. To obtain the nuclear donor, the fetal fibroblast cell of Bama (BM) pig were transfected with GHR-knockout vector in G418 selection medium. We collected 173 cell for further positive identification which showed that 46.2% (78/173) of the clones were GHR-knockout cell strains. We chose one bi-allelic knockout cell strain as nuclear donor to produce reconstructed embryos by HMC. It was shown that the blastocyst rate was 43.5% at the 6(th) day in vitro, then 654 HMC-blastocysts were transplanted to uterus of six recipient sows. Finally, a total of 10 live offspring were delivered including 7 bi-allelic knockout piglets. Fibroblasts were obtained from ear biopsies for GHR knockout detection. The body weight of the piglets was measured consecutively, and it was found that the GHR(-)(/)(-) pigs were only 50% smaller than that of the controls at the 20(th) week. In conclusion, our results indicate that TALENs and HMC technology can rapidly and efficiently produce knockout animals for agricultural and biomedical research.


Li F.,BGI Shenzhen | Li F.,ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding | Li Y.,BGI Shenzhen | Li Y.,ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding | And 18 more authors.
Transgenic Research | Year: 2015

Growth hormone (GH) is an anabolic mitogen with widespread influence on cellular growth and differentiation as well as on glucose and lipid metabolism. GH binding to the growth hormone receptor (GHR) on hepatocytes prompts expression of insulin growth factor I (IGF-1) involved in nutritionally induced compensatory hyperplasia of pancreatic β-cell islets and insulin release. A prolonged hyperactivity of the IGF-1/insulin axis in the face of insulinotropic nutrition, on the other hand, can lead to collapse of the pancreatic islets and glucose intolerance. Individuals with Laron syndrome carry mutations in the GHR gene resulting in severe congenital IGF-1 deficiency and elevated GH serum levels leading to short stature as well as perturbed lipid and glucose metabolism. However, these individuals enjoy a reduced prevalence of acne, cancer and possibly diabetes. Minipigs have become important biomedical models for human conditions due to similarities in organ anatomy, physiology, and metabolism relative to humans. The purpose of this study was to generate transgenic Wuzhishan minipigs by handmade cloning with impaired systemic GHR activity and assess their growth profile and glucose metabolism. Transgenic minipigs featuring overexpression of a dominant-negative porcine GHR (GHRdm) presented postnatal growth retardation and proportionate dwarfism. Molecular changes included elevated GH serum levels and mild hyperglycemia. We believe that this model may prove valuable in the study of GH functions in relation to cancer, diabetes and longevity. © 2015, Springer International Publishing Switzerland.


Liu H.,BGI Shenzhen | Liu H.,ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding | Li Y.,BGI Shenzhen | Li Y.,ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding | And 24 more authors.
PLoS ONE | Year: 2013

Minipigs have become important biomedical models for human ailments due to similarities in organ anatomy, physiology, and circadian rhythms relative to humans. The homeostasis of circadian rhythms in both central and peripheral tissues is pivotal for numerous biological processes. Hence, biological rhythm disorders may contribute to the onset of cancers and metabolic disorders including obesity and type II diabetes, amongst others. A tight regulation of circadian clock effectors ensures a rhythmic expression profile of output genes which, depending on cell type, constitute about 3-20% of the transcribed mammalian genome. Central to this system is the negative regulator protein Cryptochrome 1 (CRY1) of which the dysfunction or absence has been linked to the pathogenesis of rhythm disorders. In this study, we generated transgenic Bama-minipigs featuring expression of the Cys414-Ala antimorphic human Cryptochrome 1 mutant (hCRY1AP). Using transgenic donor fibroblasts as nuclear donors, the method of handmade cloning (HMC) was used to produce reconstructed embryos, subsequently transferred to surrogate sows. A total of 23 viable piglets were delivered. All were transgenic and seemingly healthy. However, two pigs with high transgene expression succumbed during the first two months. Molecular analyzes in epidermal fibroblasts demonstrated disturbances to the expression profile of core circadian clock genes and elevated expression of the proinflammatory cytokines IL-6 and TNF-α, known to be risk factors in cancer and metabolic disorders. © 2013 Liu et al.


Liu C.,BGI Shenzhen | Liu C.,ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding | Xiao L.,BGI Shenzhen | Xiao L.,ShenZhen Engineering Laboratory for Genomics Assisted Animal Breeding | And 19 more authors.
Transgenic Research | Year: 2015

The angiotensin I converting enzyme 2 (ACE2) is a key factor in the maintenance of intestinal homeostasis. Dysregulation of homeostasis can lead to inflammation of the colon (colitis), which can cause life-threatening enfeeblement or even cancer. Animal models are valuable surrogates in deciphering the pathology behind such human conditions and for screening of putative therapeutic targets or treatment paradigms. However, development of disease models can be time-consuming and technical demanding, which might hamper their application-value. In this study, we genetically disrupted the mouse Ace2 gene by direct injection of in vitro transcribed mRNA coding for transcription activator-like effector nucleases (TALENs) into the cytoplasm of outbred Kunming mouse zygotes. Consequently, somatic mutations were induced with an efficiency of 57 %, of which 39 % were frameshift mutations. Moreover, all modifications were stably transferred during germline transmission. In Ace2-knockout male mice (Ace2−/y), we observed severe chemical induced colitis, characterized by considerable weight loss, diarrhea and a shortened colon length. Histologically, Ace2 mutations resulted in the infiltration of leukocytes and the overt damage of the intestinal mucosal barrier. In addition, we detected an increased expression of inflammatory cytokines in the colon tissue of Ace2−/y mice. Collectively, the data indicate that high targeting efficiency and heritability can be achieved in an outbred mouse model by zygote injection of TALEN mRNA. Furthermore, the generated Ace2−/y mice display phenotypic traits reminiscent of colitis and we anticipate that such mice can be of value in studies of the intestinal microbiome or fecal transplantation. © 2014, Springer International Publishing Switzerland.

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