Transgenesis and Genome Editing in Poultry

The transgenic approach and precise editing of specific loci in the genome have diverse practical uses in animal biotechnology. Recent advances in genome-editing technology, including clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) have helped to generate highly valuable and quality-improved poultry. The production of transgenic and genome-edited birds mainly depends on primordial germ cells (PGCs), which are the progenitor cells of gametes, due to the unique system that is quite different from the mammalian system. This chapter introduces the basic physiology of avian PGCs and the latest PGC-mediated methodologies in transgenesis and genome editing of birds. Based on these techniques, future applications of precisely genome-modulated poultry are discussed to provide opportunities and benefits for humans

The avian-specific small heat shock protein HSP25 is a constitutive protector against environmental stresses during blastoderm dormancy

ArticleScientific Reports. 6: 36704 (2016).journal articl

A scheduling algorithm for multiport memory minimization in datapath synthesis

Abstract- In this paper, we present a new scheduling algorithms that generates area-efficient register transfer level datapaths with multiport memories. The proposed scheduling algorithm assigns an operation to a specific control step such that maximal sharing of functional units can be achieved with minimal number of memory ports, while satisfying given constraints. We propose a measure of multiport memory cost, MAV (Multiple Access Variable) which is defined as a variable accessed at several control steps, and overall memory cost is reduced by equally distributing the MAVs throughout all the control steps. When compared with previous approaches for several benchmarks available from the literature, the proposed algorithm generates the datapaths with less memory modules and interconnection structures by reflecting the memory cost in the scheduling process

5′-Triphosphate-RNA-independent activation of RIG-I via RNA aptamer with enhanced antiviral activity

RIG-I is a cytosolic receptor for non-self RNA that mediates immune responses against viral infections through IFNα/β production. In an attempt to identify novel tools that modulate IFNα/β production, we used SELEX technology to screen RNA aptamers that specifically target RIG-I protein. Most of the selected RIG-I aptamers contained polyU motifs in the second half regions that played critical roles in the activation of RIG-I-mediated IFNβ production. Unlike other known ligands, RIG-I aptamer bound and activated RIG-I in a 5′-triphosphate-independent manner. The helicase and RD domain of RIG-I were used for aptamer binding, but intact RIG-I protein was required to exert aptamer-mediated signaling activation. Furthermore, replication of NDV, VSV and influenza virus in infected host cells was efficiently blocked by pre- or post-treatment with RIG-I aptamer. Based on these data, we propose that RIG-I aptamer has strong potential to be an antiviral agent that specifically boosts the RIG-I-dependent signaling cascade

Transcription Factor Sp1 Is Involved in Expressional Regulation of Coxsackie and Adenovirus Receptor in Cancer Cells

Coxsackie and adenovirus receptor (CAR) was first known as a virus receptor. Recently, it is also known to have tumor suppressive activity such as inhibition of cell proliferation, migration, and invasion. It is important to understand how CAR expression can be regulated in cancers. Based on an existence of putative Sp1 binding site within CAR promoter, we investigated whether indeed Sp1 is involved in the regulation of CAR expression. We observed that deletion or mutation of Sp1 binding motif (−503/−498) prominently impaired the Sp1 binding affinity and activity of CAR promoter. Histone deacetylase inhibitor (TSA) treatment enhanced recruitment of Sp1 to the CAR promoter in ChIP assay. Meanwhile, Sp1 binding inhibitor suppressed the recruitment. Exogenous expression of wild-type Sp1 increased CAR expression in CAR-negative cells; meanwhile, dominant negative Sp1 decreased the CAR expression in CAR-positive cells. These results indicate that Sp1 is involved in regulation of CAR expression

Validating of the pre-clinical mouse model for metastatic breast cancer to the mandible

Metastatic breast carcinoma has a great tendency to spread to the mandible. It is concomitantly associated with bone destruction, food intake disorder, and a poorer prognosis. Appropriate animal models need to be developed for a better understanding of the mechanisms underlying the metastatic process of breast cancer cells to mandible and to test the effects of potential lead compounds. Here, we assessed the metastasis model of intracardiac injection using luciferase-transfected metastatic breast cancer cells (MDA-MB-231Luc+) by determining the incidences of metastasis, mCT images, and histopathological results. A high bioluminescence signal mainly detected mandibular lesions with less frequent distal femora and proximal tibiae lesions. Extensive mandibular bone destruction occurred in nude mice grafted with metastatic breast cancer cells. This type of animal model might be a useful tool in assessing therapeutic implications and the efficacy of anti-cancer drugs for osteolytic cancers

Crystal structure of Cmr5 from Pyrococcus furiosus and its functional implications

AbstractThe bacterial acquired immune system consists of clustered regularly interspaced short palindromic repeats (CRISPRs) and CRIPSR-associated (Cas) genes, which include Cas-module repeat-associated mysterious proteins (Cmr). The six Cmr proteins of Pyrococcus furiosus (pfCmr1–pfCmr6) form a Cmr effector complex that functions against exogenous nucleic acid. Among the Cmr proteins, the role of pfCmr5 and its involvement in the complex’s cleavage activity have been obscure. The elucidated pfCmr5 structure has two inserted α-helices compared with the other trimeric Cmr5 structure. However, pfCmr5 exists as a monomeric protein both in the crystalline state and in solution. In vitro assays indicate that pfCmr5 interacts with pfCmr4. These structural and biophysical data might help in understanding the complicated and ill-characterized Cmr effector complex.Structured summary of protein interactionspfCmr4 and pfCmr5 bind by molecular sieving (View interaction)pfCmr4 and pfCmr4 bind by molecular sieving (View interaction)pfCmr5 and pfCmr4 bind by ion exchange chromatography (View interaction

Hepatitis C Virus NS5B Protein Is a Membrane-Associated Phosphoprotein with a Predominantly Perinuclear Localization

AbstractHepatitis C virus NS5B protein is an RNA-dependent RNA polymerase. To investigate the properties and function of this protein, we have expressed the NS5B protein in insect and mammalian cells. NS5B was found to be present as fine speckles in the cytoplasm, particularly concentrated in the perinuclear region, suggesting its association with the nuclear membrane, the endoplasmic reticulum, or the Golgi complex. This conclusion was supported by the biochemical demonstration that NS5B was associated with the membranes in the cells. Furthermore, it was shown that NS5B protein is a phosphoprotein. These properties may be related to its function as an RNA polymerase

The dynamic development of germ cells during chicken embryogenesis

ArticlePoultry Science. 97(2): 650-657. (2018)journal articl