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https://www.readbyqxmd.com/read/28806146/three-dimensional-bioprinting-of-polycaprolactone-reinforced-gene-activated-bioinks-for-bone-tissue-engineering
#1
Gráinne M Cunniffe, Tomas Gonzalez-Fernandez, Andrew Daly, Binulal N Sathy, Oju Jeon, Eben Alsberg, Daniel J Kelly
Regeneration of complex bone defects remains a significant clinical challenge. Multi-tool biofabrication has permitted the combination of various biomaterials to create multifaceted composites with tailorable mechanical properties and spatially controlled biological function. In this study we sought to use bioprinting to engineer nonviral gene activated constructs reinforced by polymeric micro-filaments. A gene activated bioink was developed using RGD-γ-irradiated alginate and nano-hydroxyapatite (nHA) complexed to plasmid DNA (pDNA)...
August 14, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28805373/inefficient-ribosomal-skipping-enables-simultaneous-secretion-and-display-of-proteins-in-saccharomyces-cerevisiae
#2
Carlos A Cruz-Teran, Karthik Tiruthani, Adam Mischler, Balaji M Rao
The need for recombinant expression of soluble protein slows the validation of engineered proteins isolated from combinatorial libraries and limits the number of protein variants evaluated. To overcome this bottleneck, we describe a system for simultaneous cell surface display and soluble secretion of proteins in Saccharomyces cerevisiae based on inefficient ribosomal skipping. Ribosomal skipping mediated by "self-cleaving" 2A peptides produces two proteins from a single open reading frame. Incorporation of the F2A peptide sequence-with ∼50% efficiency of ribosomal skipping-between the protein of interest and the yeast cell wall protein Aga2 results in simultaneous expression of both the solubly secreted protein and the protein-Aga2 fusion that is tethered to the yeast cell surface...
August 14, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28804866/virally-vectored-vaccine-delivery-medical-needs-mechanisms-advantages-and-challenges
#3
Daniel D Pinschewer
Vaccines represent one of the most successful chapters in the history of medicine. Over the past decades, the advent of recombinant cDNA technology has enabled the biomedical community to genetically engineer viruses for vaccine delivery purposes. As a starting point, this review evaluates the unmet medical needs, which drive scientists and industry to exploit such fundamentally new technology for human vaccination. The author discusses the molecular functioning, production and safety profile of replication-competent and -deficient viral vector systems, representing two fundamentally distinct classes of "genetic vaccines"...
August 14, 2017: Swiss Medical Weekly
https://www.readbyqxmd.com/read/28804565/thrombospondin-1-inhibits-ossification-of-tissue-engineered-cartilage-constructed-by-adscs
#4
Aiguo Xie, Jixin Xue, Gan Shen, Lanjun Nie
Cartilage tissue engineering provides a new method in the treatment of cartilage defects, and adipose derived stem cells seem to be an ideal seed cell in cartilage tissue engineering because of its characteristics. However, ossification after in vivo implantation of tissue engineered cartilage remains a challenge. Thrombospondin-1 which has been reported to have an inhibitory effect on angiogenesis, may play an important role in inhibiting the ossification of tissue engineered cartilage constructed by adipose derived stem cells...
2017: American Journal of Translational Research
https://www.readbyqxmd.com/read/28803776/mechanism-of-enzyme-repair-by-the-aaa-chaperone-rubisco-activase
#5
Javaid Y Bhat, Goran Miličić, Gabriel Thieulin-Pardo, Andreas Bracher, Andrew Maxwell, Susanne Ciniawsky, Oliver Mueller-Cajar, John R Engen, F Ulrich Hartl, Petra Wendler, Manajit Hayer-Hartl
How AAA+ chaperones conformationally remodel specific target proteins in an ATP-dependent manner is not well understood. Here, we investigated the mechanism of the AAA+ protein Rubisco activase (Rca) in metabolic repair of the photosynthetic enzyme Rubisco, a complex of eight large (RbcL) and eight small (RbcS) subunits containing eight catalytic sites. Rubisco is prone to inhibition by tight-binding sugar phosphates, whose removal is catalyzed by Rca. We engineered a stable Rca hexamer ring and analyzed its functional interaction with Rubisco...
July 20, 2017: Molecular Cell
https://www.readbyqxmd.com/read/28803472/blue-light-switchable-bacterial-adhesion-as-a-key-step-towards-the-design-of-biofilms
#6
Fei Chen, Seraphine Valeska Wegner
The control of where and when bacterial adhere to a substrate is a key step towards controlling the formation and organization in biofilms. This study shows how we engineer bacteria to adhere specifically to substrates with high spatial and temporal control under blue light, but not in the dark, by using photoswitchable interaction between nMag and pMag proteins. For this, we express pMag proteins on the surface of E.coli so that the bacteria can adhere to substrates with immobilized nMag protein under blue light...
August 14, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28803466/deciphering-the-fluorine-code-the-many-hats-fluorine-wears-in-a-protein-environment
#7
Allison Ann Berger, Jan-Stefan Völler, Nediljko Budisa, Beate Koksch
Deciphering the fluorine code is how we describe not only the focus of this Account, but also the systematic approach to studying the impact of fluorine's incorporation on the properties of peptides and proteins used by our groups and others. The introduction of fluorine has been shown to impart favorable, but seldom predictable, properties to peptides and proteins, but up until about two decades ago the outcomes of fluorine modification of peptides and proteins were largely left to chance. Driven by the motivation to extend the application of the unique properties of the element fluorine from medicinal and agro chemistry to peptide and protein engineering we have established extensive research programs that enable the systematic investigation of effects that accompany the introduction of fluorine into this class of biopolymers...
August 12, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28803236/enhancement-of-bone-healing-by-local-administration-of-carbon-nanotubes-functionalized-with-sodium-hyaluronate-in-rat-tibiae
#8
Vanessa B Andrade, Marcos A Sá, Renato M Mendes, Paulo A Martins-Júnior, Gerluza A B Silva, Bruna R Sousa, Marcelo V Caliari, Erick S Ávila, Luiz Orlando Ladeira, Rodrigo R Resende, Anderson J Ferreira
It has been reported that carbon nanotubes (CNTs) serve as nucleation sites for the deposition of bone matrix and cell proliferation. Here, we evaluated the effects of multi-walled CNTs (MWCNTs) on bone repair of rat tibiae. Furthermore, because sodium hyaluronate (HY) accelerates bone restoration, we associated CNTs with HY (HY-MWCNTs) in an attempt to boost bone repair. The bone defect was created by a 1.6-mm-diameter drill. After 7 and 14 days, tibiae were processed for histological and morphometric analyses...
August 12, 2017: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/28802102/pnipaam-co-jeffamine-%C3%A2-pnj-scaffolds-as-in%C3%A2-vitro-models-for-niche-enrichment-of-glioblastoma-stem-like-cells
#9
John M Heffernan, James B McNamara, Sabine Borwege, Brent L Vernon, Nader Sanai, Shwetal Mehta, Rachael W Sirianni
Glioblastoma (GBM) is the most common adult primary brain tumor, and the 5-year survival rate is less than 5%. GBM malignancy is driven in part by a population of GBM stem-like cells (GSCs) that exhibit indefinite self-renewal capacity, multipotent differentiation, expression of neural stem cell markers, and resistance to conventional treatments. GSCs are enriched in specialized niche microenvironments that regulate stem phenotypes and support GSC radioresistance. Therefore, identifying GSC-niche interactions that regulate stem phenotypes may present a unique target for disrupting the maintenance and persistence of this treatment resistant population...
May 6, 2017: Biomaterials
https://www.readbyqxmd.com/read/28801913/design-of-synthetic-promoters-for-gene-circuits-in-mammalian-cells
#10
Pratik Saxena, Daniel Bojar, Martin Fussenegger
Synthetic biology, the synthesis of engineering and biology, has rapidly matured and has dramatically increased the complexity of artificial gene circuits in recent years. The deployment of intricate synthetic gene circuits in mammalian cells requires the establishment of very precise and orthogonal control of transgene expression. In this chapter, we describe methods of modulating the expression of transgenes at the transcriptional level. Using cAMP-response element-binding protein (CREB)-dependent promoters as examples, a tool for the precise tuning of gene expression by using different core promoters and by varying the binding affinity of transcription factor operator sites is described...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28800110/response-of-dermal-fibroblasts-to-biochemical-and-physical-cues-in-aligned-polycaprolactone-silk-fibroin-nanofiber-scaffolds-for-application-in-tendon-tissue-engineering
#11
Chih-Hao Chen, Shih-Hsien Chen, Chang-Yi Kuo, Meng-Lun Li, Jyh-Ping Chen
Silk fibroin (SF) and fiber alignment were introduced into polycaprolactone (PCL)-based electrospun nanofibers as chemical and physical cues for tendon tissue engineering applications. The physicochemical properties of random PCL (RP) nanofibers, random PCL/SF (RPSF) nanofibers and aligned PCL/SF (APSF) nanofibers were characterized for fiber orientation and SF blending effects. An in vitro cell culture with rabbit dermal fibroblasts (RDFBs) on nanofibers indicated that SF promotes cell proliferation to a higher extent than fiber alignment...
August 11, 2017: Nanomaterials
https://www.readbyqxmd.com/read/28799089/searching-for-the-pareto-frontier-in-multi-objective-protein-design
#12
REVIEW
Vikas Nanda, Sandeep V Belure, Ofer M Shir
The goal of protein engineering and design is to identify sequences that adopt three-dimensional structures of desired function. Often, this is treated as a single-objective optimization problem, identifying the sequence-structure solution with the lowest computed free energy of folding. However, many design problems are multi-state, multi-specificity, or otherwise require concurrent optimization of multiple objectives. There may be tradeoffs among objectives, where improving one feature requires compromising another...
August 10, 2017: Biophysical Reviews
https://www.readbyqxmd.com/read/28798063/alignment-of-the-protein-substrate-hairpin-along-the-seca-two-helix-finger-primes-protein-transport-in-escherichia-coli
#13
Qi Zhang, Sudipta Lahiri, Tithi Banerjee, Zhongmou Sun, Donald Oliver, Ishita Mukerji
A conserved hairpin-like structure comprised of a signal peptide and early mature region initiates protein transport across the SecY or Sec61α channel in Bacteria or Archaea and Eukarya, respectively. When and how this initiator substrate hairpin forms remains a mystery. Here, we have used the bacterial SecA ATPase motor protein and SecYEG channel complex to address this question. Engineering of a functional miniprotein substrate onto the end of SecA allowed us to efficiently form ternary complexes with SecYEG for spectroscopic studies...
August 10, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28796529/designed-ankyrin-repeat-proteins-as-her2-targeting-domains-in-chimeric-antigen-receptor-engineered-t-cells
#14
Elizabeth Siegler, Si Li, Yu Jeong Kim, Pin Wang
Chimeric antigen receptor (CAR) engineering is a branch of cancer immunotherapy that equips immune cells to target tumor antigens expressed on the cell surface using antibody-derived single-chain variable fragments (scFvs). However, other antibody mimetics, such as designed ankyrin repeat proteins (DARPins), can also serve as antigen-binding domains in CARs. This study shows that CAR-engineered T (CAR-T) cells utilizing Her2-targeting DARPins G3 and 929 can target human epidermal growth factor receptor 2 (Her2)-overexpressing cancer cells as effectively as CAR-T cells with the scFv 4D5 in vitro, and G3 CAR-T cells can slow or eliminate tumor growth in vivo as effectively as 4D5 CAR-T cells...
June 22, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28796297/transfer-of-tomato-immune-receptor-ve1-confers-ave1-dependent-verticillium-resistance-in-tobacco-and-cotton
#15
Yin Song, Linlin Liu, Yidong Wang, Dirk-Jan Valkenburg, Xianlong Zhang, Longfu Zhu, Bart P H J Thomma
Verticillium wilts caused by soil-borne fungal species of the Verticillium genus are economically important plant diseases that affect a wide range of host plants, and are notoriously difficult to combat. Perception of pathogen(-induced) ligands by plant immune receptors is a key component of plant innate immunity. In tomato, race-specific resistance to Verticillium wilt is governed by the cell surface-localized immune receptor Ve1 through recognition of the effector protein Ave1 that is secreted by race 1 strains of Verticillium spp...
August 10, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28796233/intracellular-directed-evolution-of-proteins-from-combinatorial-libraries-based-on-conditional-phage-replication
#16
Andreas K Brödel, Alfonso Jaramillo, Mark Isalan
Directed evolution is a powerful tool to improve the characteristics of biomolecules. Here we present a protocol for the intracellular evolution of proteins with distinct differences and advantages in comparison with established techniques. These include the ability to select for a particular function from a library of protein variants inside cells, minimizing undesired coevolution and propagation of nonfunctional library members, as well as allowing positive and negative selection logics using basally active promoters...
September 2017: Nature Protocols
https://www.readbyqxmd.com/read/28796191/smart-carriers-and-nanohealers-a-nanomedical-insight-on-natural-polymers
#17
REVIEW
Sreejith Raveendran, Ankit K Rochani, Toru Maekawa, D Sakthi Kumar
Biodegradable polymers are popularly being used in an increasing number of fields in the past few decades. The popularity and favorability of these materials are due to their remarkable properties, enabling a wide range of applications and market requirements to be met. Polymer biodegradable systems are a promising arena of research for targeted and site-specific controlled drug delivery, for developing artificial limbs, 3D porous scaffolds for cellular regeneration or tissue engineering and biosensing applications...
August 10, 2017: Materials
https://www.readbyqxmd.com/read/28795766/computerized-implementation-of-higher-order-electron-correlation-methods-and-their-linear-scaling-divide-and-conquer-extensions
#18
Masahiko Nakano, Takeshi Yoshikawa, So Hirata, Junji Seino, Hiromi Nakai
We have implemented a linear-scaling divide-and-conquer (DC)-based higher-order coupled-cluster (CC) and Møller-Plesset perturbation theories (MPPT) as well as their combinations automatically by means of the tensor contraction engine, which is a computerized symbolic algebra system. The DC-based energy expressions of the standard CC and MPPT methods and the CC methods augmented with a perturbation correction were proposed for up to high excitation orders [e.g., CCSDTQ, MP4, and CCSD(2)TQ ]. The numerical assessment for hydrogen halide chains, polyene chains, and first coordination sphere (C1) model of photoactive yellow protein has revealed that the DC-based correlation methods provide reliable correlation energies with significantly less computational cost than that of the conventional implementations...
August 10, 2017: Journal of Computational Chemistry
https://www.readbyqxmd.com/read/28795725/rational-design-of-artificial-dye-decolorizing-peroxidases-using-myoglobin-by-engineering-tyr-trp-in-the-heme-center
#19
Le-Le Li, Hong Yuan, Fei Liao, Bo He, Shu-Qin Gao, Ge-Bo Wen, Xiangshi Tan, Ying-Wu Lin
The rational design of metalloenzymes provides advantages not only for illustrating the structure and function relationship of native enzymes, but also for creating functional artificial enzymes comparable to native enzymes. Dye-decolorizing peroxidases (DyPs) are a new family of heme peroxidases and have received much attention recently. Inspired by the structural features of native DyPs with multiple Tyr and Trp residues, we herein aimed to design functional artificial DyPs using myoglobin (Mb), an O2 carrier as a protein scaffold, by further introduction of Tyr/Trp into the secondary sphere of the heme center in the F43Y Mb mutant...
August 10, 2017: Dalton Transactions: An International Journal of Inorganic Chemistry
https://www.readbyqxmd.com/read/28795570/rapid-and-scalable-preparation-of-bacterial-lysates-for-cell-free-gene-expression
#20
Andriy Didovyk, Taishi Tonooka, Lev Tsimring, Jeff Hasty
Cell-free gene expression systems are emerging as an important platform for a diverse range of synthetic biology and biotechnology applications, including production of robust field-ready biosensors. Here, we combine programmed cellular autolysis with a freeze-thaw or freeze-dry cycle to create a practical, reproducible, and a labor- and cost-effective approach for rapid production of bacterial lysates for cell-free gene expression. Using this method, robust and highly active bacterial cell lysates can be produced without specialized equipment at a wide range of scales, making cell-free gene expression easily and broadly accessible...
August 10, 2017: ACS Synthetic Biology
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