Abstract:
:Increases in our understanding of gene function have greatly expanded the repertoire of possible genetic interventions at our disposal with the consequence that many genetic engineering applications require multiple manipulations in which target genes can be both overexpressed and silenced in a simple and co-ordinated manner. Using synthetic introns as a source of encoding short-interfering RNA (siRNA), we demonstrate that it is possible to simultaneously express both a transgene and siRNA from a single polymerase (Pol) II promoter. By encoding siRNA as an intron between two protein domains requiring successful splicing for functionality, it was possible to demonstrate that splicing was occurring, that the coding genes (exonic transgenes) resulted in functional protein, and that the spliced siRNA-containing lariat was capable of modulating expression of a separate target gene. We subsequently extended this concept to develop pTRIDENT-based multi-cistronic vectors that were capable of co-ordinated expression of up to three siRNAs and three transgenes off a single genetic platform. Such multi-gene engineering technology, enabling concomitant transgene overexpression and target gene knockdown, should be useful for therapeutic, biopharmaceutical production, and basic research applications.
journal_name
Biotechnol Bioengjournal_title
Biotechnology and bioengineeringauthors
Greber D,Fussenegger Mdoi
10.1002/bit.21303subject
Has Abstractpub_date
2007-04-01 00:00:00pages
821-34issue
5eissn
0006-3592issn
1097-0290journal_volume
96pub_type
杂志文章abstract::During lignocellulosic ethanol fermentation, yeasts are exposed to various lignocellulose-derived inhibitors, which disrupt the efficiency of hexose and pentose co-fermentation. To understand the metabolic response of fermentation microbes to these inhibitors, a comparative metabolomic investigation was performed on a...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.24992
更新日期:2014-01-01 00:00:00
abstract::Osteochondral tissue engineering aims to regenerate functional tissue-mimicking physiological properties of injured cartilage and its subchondral bone. Given the distinct structural and biochemical difference between bone and cartilage, bilayered scaffolds, and bioreactors are commonly employed. We present an osteocho...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.27127
更新日期:2019-11-01 00:00:00
abstract::This article describes a method for the chemical immobilization of concanavalin A (Con A) on the inside wall of a single hollow cellulose fiber for use in glucose affinity sensor. Periodate oxidation of cellulose fiber followed by a spacer for Con A attachment was deemed to be the most optimal procedure for achieving ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260280213
更新日期:1986-02-01 00:00:00
abstract::The growth and product formation of a Savinase-producing Bacillus clausii were investigated in high-cell-density fed-batch cultivations with both linear and exponential feed profiles. The highest specific productivity of Savinase was observed shortly after the end of the initial batch phase for all feed profiles appli...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10675
更新日期:2003-08-05 00:00:00
abstract::We designed a bioartificial liver support system in which encapsulated multicellular spheroids of rat hepatocytes were utilized as a bioreactor in a hollow fiber cartridge. The spheroids, formed in a positively charged polystyrene dish that contained hormonally defined medium, were encapsulated into microdroplets of a...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(SICI)1097-0290(19960520)50:4<416::AID-BIT
更新日期:1996-05-20 00:00:00
abstract::In the present investigation molecular sieve has been found to be a suitable carrier for the immobilization of enzymes. From the present study it may be specifically found that trypsin and pappain can be immobilized by molecular sieve type 4a following a very simple techniqure. The immobilized enzyme can be used both ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260190902
更新日期:1977-09-01 00:00:00
abstract::Mechanical forces regulate the structure and function of many tissues in vivo; recent results indicate that the mechanical environment can decisively influence the development of engineered tissues cultured in vitro. To investigate the effects of the hydrodynamic environment on tissue-engineered cartilage, primary bov...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/1097-0290(20000220)72:4<402::aid-bit1002>3
更新日期:2001-02-20 00:00:00
abstract::The yeast Pachysolen tannophilus was entrapped in calcium alginate beads to ferment D-xylose on a continous basis in the presence of high cell densities. Experimental operating variables included the feed D-xylose concentration, the dilution rate, and the fermentor biomass concentration. Under favorable operating cond...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260241010
更新日期:1982-10-01 00:00:00
abstract::Regenerative medicine has emerged as an important discipline that aims to repair injury or replace damaged tissues or organs by introducing living cells or functioning tissues. Successful regenerative medicine strategies will likely depend upon a simultaneous optimization strategy for the design of biomaterials, cell-...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.26452
更新日期:2018-01-01 00:00:00
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journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22937
更新日期:2011-02-01 00:00:00
abstract::Alkyl beta-D-glucosides were synthesized from D-glucose and alcohols by reverse hydrolysis using the commercially available almond beta-D-glucosidase in 9:1 (v/v) acetonitrile-water medium. The main characteristics of this enzyme-catalyzed glucosylation were established by using 2-hydroxybenzyl alcohol. The reaction i...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260460204
更新日期:1995-04-20 00:00:00
abstract::Model experiments were performed with starved yeast (Saccharomyces cerevisiae) cultures in a batch reactor in order to develop a better understanding of NAD(P)H and flavin culture fluorescence. Fluorescence was monitored during aerobic-anaerobic-aerobic transitions and ethanol and glucose substrate addition experiment...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260340510
更新日期:1989-08-20 00:00:00
abstract::We investigated the enzymic activity of bilirubin oxidase when it is suspended as a lyophilized powder in a low-water system. The enzyme required buffer salts and a source of water to show activity. This study investigated the complete range of water thermodynamic activity (a(w)) by combining the use of salt hydrates ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10103
更新日期:2002-03-20 00:00:00
abstract::The thermophilic fungus Sporotrichum cellulophilum was immobilized with nonwoven materials for cellulase production. The cellulose powder concentration in the medium was an important factor controlling cellulase production. When the cellulose powder concentration in the nonwoven materials was more than 4%, cellulase p...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260280814
更新日期:1986-08-01 00:00:00
abstract::Lignocellulosic biofuels and chemicals have great potential to reduce our dependence on fossil fuels and mitigate air pollution by cutting down on greenhouse gas emissions. Chemical, thermal, and enzymatic processes are used to release the sugars from the lignocellulosic biomass for conversion to biofuels. These proce...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章,评审
doi:10.1002/bit.27639
更新日期:2020-11-26 00:00:00
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journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.25712
更新日期:2016-02-01 00:00:00
abstract::Engineering protein expression in vitro or in vivo is usually straightforward for single genes, but remains challenging for multiple genes because of the requirement of coordinated control. RNA and protein overexpression strategies often exploit T7 RNA polymerase and its natural TPhi Class I terminator. However, this ...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22491
更新日期:2009-12-15 00:00:00
abstract::Hyperacute rejection following xenogeneic transplantation in primates is mediated by naturally occurring IgM antibodies, which are specifically directed to alpha-Galactosyl residues on many nonprimate mammalian cells. Current approaches to remove these anti-alphaGal IgM include plasmapheresis followed by immunoaffinit...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10555
更新日期:2003-01-20 00:00:00
abstract::Trypsin and alpha-chymotrypsin were immobilized by gelentrapment in polyacrylamide cross-linked with N,N(1)-methylenebisacrylamide. The immobilized enzymes are catalytically efficient in suspensions of reverse micelles formed in isooctane by bis(2-ethylhexyl) sodium sulfosuccinate (AOT) and water. Both entrapped enzym...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260331008
更新日期:1989-04-20 00:00:00
abstract::The bioconversion of octanoic acid into 2-heptanone by spores of Penicillium roquefortii is performed using a fed-batch technique with pH control by addition of the liquid substrate itself. The early stage of this process takes place with a high bioconversion rate and high yield. These values then decrease as a result...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260440606
更新日期:1994-09-05 00:00:00
abstract::Cellulose fibers in water form networks that give rise to an apparent yield stress, especially at high solids contents. Measuring the yield stress and correlating it with fiber concentration is important for the biomass and pulp industries. Understanding how the yield stress behaves at high solids concentrations is cr...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
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更新日期:2011-10-01 00:00:00
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journal_title:Biotechnology and bioengineering
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更新日期:2007-05-01 00:00:00
abstract::Potentiometric and amperometric measurements were made with microbial fuel cells containing E. coli or yeast as the anodic reducing agent and glucose as the oxidizable substrate. The catalytic effects of thionine and resorufin on the anode reaction were investigated. Results on the potentiometry, polarization, and cou...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260250219
更新日期:1983-02-01 00:00:00
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journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.22277
更新日期:2009-06-15 00:00:00
abstract::The glycosylation patterns of recombinant therapeutic glycoproteins can be engineered by overexpression of glycosyltransferases in the host cells used for glycoprotein production. Most prior glycosylation engineering experiments have involved constitutive expression of cloned glycosyltransferases. Here we use tetracyc...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:
更新日期:1999-12-05 00:00:00
abstract::While elastin-like polypeptides and peptides (ELPs) have been used for various stimulus-responsive applications, details of their switching remain unclear. We therefore constructed a novel series of filamentous phage particles displaying a high surface density of short ELPs. The surface display of ELPs did not disrupt...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.24854
更新日期:2013-07-01 00:00:00
abstract::Downstream processing operations are often carried out blind in the process timescale since product monitoring on-line is not common. Knowledge of the location and concentration of the product and key contaminants is complementary to other process information for process development and, if available on-line in conjun...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/(SICI)1097-0290(19970105)53:1<58::AID-BIT9
更新日期:1997-01-05 00:00:00
abstract::In order to carry out an effective enzyme reaction, the preparation of soluble-insoluble immobilized enzyme was investigated. Proteases were selected as model enzymes, and their immobilization was carried out by using an enteric coating polymer as a carrier. Among the polymers tested, methacrylic acid-methylacrylate-m...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.260290612
更新日期:1987-04-01 00:00:00
abstract::The effect of trehalose (0.5 M) on the thermal stability of cutinase in the alkaline pH range was studied. The thermal unfolding induced by increasing temperature was analyzed in the absence and in the presence of trehalose according to a two-state model (which assumes that only the folded and unfolded states of cutin...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
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更新日期:2000-12-20 00:00:00
abstract::This paper proposes a new metabolic model for acetate uptake by a mixed culture of phosphate- and glycogen-accumulating organisms (PAOs and GAOs) under anaerobic conditions. The model uses variable overall stoichiometry based on the assumption that PAOs may have the ability of using the glyoxylate pathway to produce t...
journal_title:Biotechnology and bioengineering
pub_type: 杂志文章
doi:10.1002/bit.10765
更新日期:2003-11-05 00:00:00