RepSox Increases Porcine Cloning Efficiency by Improving Pluripotency of Donor Nuclei

2019 ◽  
Vol 21 (4) ◽  
pp. 181-186 ◽  
Author(s):  
Guosong Qin ◽  
Jianguo Zhao ◽  
Jiaojiao Huang
Keyword(s):  
Cloning Efficiency

Influence of oocyte donor and embryo recipient conditions on cloning efficiency in dogs

2010 ◽  
Vol 74 (3) ◽  
pp. 473-478 ◽  
Author(s):  
M.J. Kim ◽  
H.J. Oh ◽  
J.E. Park ◽  
S.G. Hong ◽  
J.T. Kang ◽  
...  
Keyword(s):  
Cloning Efficiency ◽  
Oocyte Donor

Human κ chain expression in a λ phage vector: methods of isolating amplified cDNA affects cloning efficiency

1993 ◽  
Vol 4 (3) ◽  
pp. 104-106
Author(s):  
Konstantinos Lazaridis ◽  
Howard Dang ◽  
Norman Talal
Keyword(s):  
Cloning Efficiency

Successful cloning of coyotes through interspecies somatic cell nuclear transfer using domestic dog oocytes

2013 ◽  
Vol 25 (8) ◽  
pp. 1142 ◽  
Author(s):  
Insung Hwang ◽  
Yeon Woo Jeong ◽  
Joung Joo Kim ◽  
Hyo Jeong Lee ◽  
Mina Kang ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Canis Lupus ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Fusion Rate ◽  
Domestic Dogs ◽  
Domestic Dog ◽  
Canis Lupus Familiaris ◽  
Cloning Efficiency ◽  
Donor Cells

Interspecies somatic cell nuclear transfer (iSCNT) is an emerging assisted reproductive technology (ART) for preserving Nature’s diversity. The scarcity of oocytes from some species makes utilisation of readily available oocytes inevitable. In the present study, we describe the successful cloning of coyotes (Canis latrans) through iSCNT using oocytes from domestic dogs (Canis lupus familiaris or dingo). Transfer of 320 interspecies-reconstructed embryos into 22 domestic dog recipients resulted in six pregnancies, from which eight viable offspring were delivered. Fusion rate and cloning efficiency during iSCNT cloning of coyotes were not significantly different from those observed during intraspecies cloning of domestic dogs. Using neonatal fibroblasts as donor cells significantly improved the cloning efficiency compared with cloning using adult fibroblast donor cells (P < 0.05). The use of domestic dog oocytes in the cloning of coyotes in the present study holds promise for cloning other endangered species in the Canidae family using similar techniques. However, there are still limitations of the iSCNT technology, as demonstrated by births of morphologically abnormal coyotes and the clones’ inheritance of maternal domestic dog mitochondrial DNA.

scholarly journals Increased Cloning Efficiency by Temperature-Cycle Ligation

1996 ◽  
Vol 24 (4) ◽  
pp. 800-801 ◽  
Author(s):  
A. H. Lund ◽  
M. Duch ◽  
F. Skou Pedersen
Keyword(s):  
Temperature Cycle ◽  
Cloning Efficiency

scholarly journals Somatic Cell Nuclear Transfer Embryos Show Massive Dysregulation of Genes Involved in Transcription Pathway

2020 ◽  
Author(s):  
Chunshen Long ◽  
Hanshuang Li ◽  
Xinru Li ◽  
Yongchun Zuo
Keyword(s):  
Embryo Development ◽  
Nuclear Transfer ◽  
Normal Embryo ◽  
Cloning Efficiency ◽  
Rna Seq ◽  
Activation Of Transcription ◽  
Potential Association ◽  
Gene Regulatory ◽  
Genome Activation ◽  
Zygotic Genome

AbstractTranscription is the most fundamental molecular event that occurs with zygotic genome activation (ZGA) during embryo development. However, the potential association between transcription pathways and low cloning efficiency of nuclear transfer (NT) embryos remains elusive. Here, we integrated a series of RNA-seq data on NT embryos to deciphering the molecular barriers of NT embryo development. Comparative transcriptome analysis indicated that incomplete activation of transcription pathways functions as a barrier for NT embryos. Then, the gene regulatory network (GRN) identified that crucial factors responsible for transcription play a coordinated role in epigenome erasure and pluripotency regulation during normal embryo development. But in NT embryos, massive genes involved in transcription pathways were varying degrees of inhibition. Our study therefore provides new insights into understanding the barriers to NT embryo reprogramming.

scholarly journals Label-free cell based impedance measurements of ZnO nanoparticles—human lung cell interaction: a comparison with MTT, NR, Trypan blue and cloning efficiency assays

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Giuseppina Bozzuto ◽  
Giuseppe D’Avenio ◽  
Maria Condello ◽  
Simona Sennato ◽  
Ezio Battaglione ◽  
...  
Keyword(s):  
Trypan Blue ◽  
A549 Cells ◽  
Cell Interaction ◽  
Neutral Red ◽  
Free Cell ◽  
Mitogenic Activity ◽  
Label Free ◽  
Cloning Efficiency ◽  
Zno Nps ◽  
Experimental Conditions

Abstract Background There is a huge body of literature data on ZnOnanoparticles (ZnO NPs) toxicity. However, the reported results are seen to be increasingly discrepant, and deep comprehension of the ZnO NPs behaviour in relation to the different experimental conditions is still lacking. A recent literature overview emphasizes the screening of the ZnO NPs toxicity with more than one assay, checking the experimental reproducibility also versus time, which is a key factor for the robustness of the results. In this paper we compared high-throughput real-time measurements through Electric Cell-substrate Impedance-Sensing (ECIS®) with endpoint measurements of multiple independent assays. Results ECIS-measurements were compared with traditional cytotoxicity tests such as MTT, Neutral red, Trypan blue, and cloning efficiency assays. ECIS could follow the cell behavior continuously and noninvasively for days, so that certain long-term characteristics of cell proliferation under treatment with ZnO NPs were accessible. This was particularly important in the case of pro-mitogenic activity exerted by low-dose ZnO NPs, an effect not revealed by endpoint independent assays. This result opens new worrisome questions about the potential mitogenic activity exerted by ZnO NPs, or more generally by NPs, on transformed cells. Of importance, impedance curve trends (morphology) allowed to discriminate between different cell death mechanisms (apoptosis vs autophagy) in the absence of specific reagents, as confirmed by cell structural and functional studies by high-resolution microscopy. This could be advantageous in terms of costs and time spent. ZnO NPs-exposed A549 cells showed an unusual pattern of actin and tubulin distribution which might trigger mitotic aberrations leading to genomic instability. Conclusions ZnO NPs toxicity can be determined not only by the intrinsic NPs characteristics, but also by the external conditions like the experimental setting, and this could account for discrepant data from different assays. ECIS has the potential to recapitulate the needs required in the evaluation of nanomaterials by contributing to the reliability of cytotoxicity tests. Moreover, it can overcome some false results and discrepancies in the results obtained by endpoint measurements. Finally, we strongly recommend the comparison of cytotoxicity tests (ECIS, MTT, Trypan Blue, Cloning efficiency) with the ultrastructural cell pathology studies. Graphic Abstract

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