Abstract:
:In this study, we compared the gene targeting efficiencies of two rAAV-BRCA1 KO targeting constructs in Yucatan and Göttingen minipig fibroblasts. The homology arms of the constructs consisted exclusively of exonic sequences amplified by PCR from Yucatan genomic DNA. The sequences were identical to those of the reference porcine genome of a Duroc sow (Ensembl Susscrofa 9) and the BRCA1 gene of the Landrace breed (NCBI acc. no. AB271921). Surprisingly, we found that the very efficient gene targeting observed for Yucatan fibroblasts (35% targeting efficiency) was completely absent using either of the two constructs in Göttingen fibroblasts. Sequencing of the relevant BRCA1 exon 11 region (~2 kb) in the Göttingen minipig revealed three single nucleotide differences in the sequence targeted by the left homology arm of the construct (0.3% of the bases) and three or seven in the two right homology regions (0.3 or 0.7% of the bases, respectively). Construction of a novel rAAV-BRCA1 targeting vector based on the Göttingen genomic DNA sequence re-established gene targeting although the efficiency was somewhat lower than that observed for Yucatan fibroblasts. These BRCA1 KO Göttingen fibroblast clones have been used as nuclear donor cells for somatic cell nuclear transfer to generate a Göttingen BRCA1 KO pig model as previously done with the Yucatan breed. The present study illustrates that even a few mismatches present in the homology arms of an efficient rAAV-targeting construct can completely abolish gene targeting by homologous recombination emphasizing the importance of using isogenic DNA even for creating targeting constructs consisting of exon sequences only.
journal_name
Transgenic Resjournal_title
Transgenic researchauthors
Luo Y,Bolund L,Sørensen CBdoi
10.1007/s11248-011-9563-1subject
Has Abstractpub_date
2012-06-01 00:00:00pages
671-6issue
3eissn
0962-8819issn
1573-9368journal_volume
21pub_type
杂志文章abstract::The development of drought tolerant plants is a high priority because the area suffering from drought is expected to increase in the future due to global warming. One strategy for the development of drought tolerance is to genetically engineer plants with transcription factors (TFs) that regulate the expression of sev...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-013-9723-6
更新日期:2014-02-01 00:00:00
abstract::It is generally believed that too high or low levels of endothelin-1 (ET-1), a strong vasoconstrictor, may be detrimental to animals. Therefore, in order to understand the in vivo function of ET-1, we used a conditional transgenic approach, Cre/loxP recombination system, to generate transgenic mice that over-express E...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1023/b:trag.0000026072.01351.10
更新日期:2004-04-01 00:00:00
abstract::Follicle-stimulating hormone (FSH) is a critical hormone regulating reproduction in mammals. Transgenic mice show that overexpression of FSH can improve female fecundity. Using a bacterial artificial chromosome (BAC) system and somatic cell nuclear transfer, we herein generated 67 Large White transgenic (TG) boars har...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-016-9963-3
更新日期:2016-10-01 00:00:00
abstract::The low-level presence (LLP) of genetically engineered (GE) seeds that have been approved in the country of origin but not the country of import presents challenges for regulators in both seed importing and exporting countries, as well as for the international seed trade and the farmers who rely on it. In addition to ...
journal_title:Transgenic research
pub_type: 杂志文章,评审
doi:10.1007/s11248-015-9899-z
更新日期:2015-10-01 00:00:00
abstract::To examine the consequences of increased apolipoprotein A-I production on cholesterol and lipoprotein metabolism, we have produced two lines of transgenic rats; one expressing moderate and one very high levels of human apolipoprotein A-I. The rats were produced by microinjection of a 13 kbp DNA fragment containing the...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/BF02528779
更新日期:1992-05-01 00:00:00
abstract::This study aims to investigate the relative plant growth and reproduction of insect-resistant and susceptible plants following the introgression of an insect-resistance Bt-transgene from Brassica napus, oilseed rape, to wild Brassica juncea. The second backcrossed generation (BC2) from a single backcross family was gr...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-014-9858-0
更新日期:2015-06-01 00:00:00
abstract::The development of animal biotechnology has led to an increase in attention to biosafety issues. Here we evaluated the impact of genetically engineered cows on the environment. The probability of horizontal gene transfer and the impact on the microbial communities in cow gut and soil were tested using three varieties ...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-010-9477-3
更新日期:2011-10-01 00:00:00
abstract::Wheat and barley are the major temperate cereals, being used for food, feed and industrial raw material. However, in all cases the quality may be limited by the amount, composition and properties of the grain storage proteins. We describe how a combination of biochemical and molecular studies has led to an understandi...
journal_title:Transgenic research
pub_type: 杂志文章,评审
doi:10.1007/BF01976021
更新日期:1994-01-01 00:00:00
abstract::To characterize the activity of the polyomavirus regulatory region, two hybrid marker genes were constructed. In the first construct, the early promoter regulates expression of the CAT gene and the late promoter regulates expression of the lacZ gene. In the second construct, the lacZ gene was placed under the control ...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/BF01980202
更新日期:1996-11-01 00:00:00
abstract::The cDNA for bovine spleen trypsin inhibitor (SI), a homologue of bovine pancreatic trypsin inhibitor (BPTI), including the natural mammalian presequence was expressed in tobacco using Agrobacterium tumefaciens-mediated transformation. Stable expression required the N-terminal targeting signal presequence although sub...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1023/a:1015210919077
更新日期:2002-04-01 00:00:00
abstract::Alpha1 tubulin gene expression is induced in the developing and regenerating CNS of vertebrates. Therefore, alpha1 tubulin gene expression may serve as a good probe for mechanisms underlying CNS development and regeneration. One approach to identify these mechanisms is to work backwards from the genome. This requires ...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1023/a:1008998832552
更新日期:2001-01-01 00:00:00
abstract::Camelina (Camelina sativa (L.) Crantz) is a re-emergent oilseed crop that is also becoming important as a model for applied projects based on studies in Arabidopsis thaliana, since the two species are closely related members of the tribe Camelineae of the Brassicaeae. Since camelina can be transformed genetically by f...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-013-9722-7
更新日期:2014-02-01 00:00:00
abstract::Rotaviruses are the main cause of infantile viral gastroenteritis worldwide leading to approximately 500,000 deaths each year mostly in the developing world. For unknown reasons, live attenuated viruses used in classical vaccine strategies were shown to be responsible for intussusception (a bowel obstruction). New str...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-005-1771-0
更新日期:2005-12-01 00:00:00
abstract::The activated cannabinoid receptor 1 (CB1R) is exclusively responsible for food intake and weight gain and regulates several pathological features associated with obesity in mammals. However, the precise role of CB1R in non-mammalian model systems is poorly understood. To investigate the functions of CB1R in zebrafish...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-012-9685-0
更新日期:2013-08-01 00:00:00
abstract::The beta-glucuronidase (GUS) gene is to date the most frequently used reporter gene in plants. Marketing of crops containing this gene requires prior evaluation of their biosafety. To aid such evaluations of the GUS gene, irrespective of the plant into which the gene has been introduced, the ecological and toxicologic...
journal_title:Transgenic research
pub_type: 杂志文章,评审
doi:10.1023/a:1008832711805
更新日期:1998-05-01 00:00:00
abstract::With the potential advent of genetically modified herbicide-resistant (GMHR) crops in the European Union, changes in patterns of herbicide use are predicted. Broad-spectrum, non-selective herbicides used with GMHR crops are expected to substitute for a set of currently used herbicides, which might alter the agro-envir...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-008-9181-8
更新日期:2008-12-01 00:00:00
abstract::Japanese morning glory, Ipomoea nil, exhibits a variety of flower colours, except yellow, reflecting the accumulation of only trace amounts of carotenoids in the petals. In a previous study, we attributed this effect to the low expression levels of carotenogenic genes in the petals, but there may be other contributing...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-017-0051-0
更新日期:2018-02-01 00:00:00
abstract::Two commonly used promoters to ubiquitously express transgenes in zebrafish are the Xenopus laevis elongation factor 1 alpha promoter (XlEef1a1) and the zebrafish histone variant H2A.F/Z (h2afv) promoter. Recently, transgenes utilizing these promoters were shown to be silenced in certain adult tissues, particularly th...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-007-9152-5
更新日期:2008-04-01 00:00:00
abstract::Agriculture productivity is severely affected by soil salinity. One possible mechanism by which plants could survive salt stress is to compartmentalize sodium ions away from the cytosol. In the present work, transgenic buckwheat plants overexpressing AtNHX1, a vacuolar Na(+)/H(+) antiporter gene from Arabidopsis thali...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-007-9085-z
更新日期:2008-02-01 00:00:00
abstract::Multiple infections of Sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV), Sweet potato virus G (SPVG) and Sweet potato mild mottle virus (SPMMV) cause a devastating synergistic disease complex of sweet potato (Ipomoea batatas Lam.) in KwaZulu-Natal, South Africa. In order to addres...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-013-9759-7
更新日期:2014-04-01 00:00:00
abstract::One of the most powerful strategies to investigate biology we have as scientists, is the ability to transfer genetic material in a controlled and deliberate manner between organisms. When applied to livestock, applications worthy of commercial venture can be devised. Although initial methods used to generate transgeni...
journal_title:Transgenic research
pub_type: 杂志文章,评审
doi:10.1007/s11248-016-9932-x
更新日期:2016-06-01 00:00:00
abstract::Polyamines, spermidine, spermine and their precursor putrescine, are ubiquitous cell components essential for normal cell growth. Increased polyamine levels and enhanced biosynthesis have been associated with malignant transformation and tumor formation, and thus, the polyamines have been considered to be a meaningful...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-013-9763-y
更新日期:2014-02-01 00:00:00
abstract::Lentiviral technology is a powerful tool for the creation of stable transgenic animals. However, uncertainties have remained whether constitutive promoters resist long-term silencing. We used concentrated HIV-1 based lentiviral vectors to create stable transgenic BALB/c mice by perivitelline injection. In our vectors ...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-009-9288-6
更新日期:2010-02-01 00:00:00
abstract::Several reports of promoters from plants, viral and artificial origin that confer high constitutive expression are known. Among these the CaMV 35S promoter is used extensively for transgene expression in plants. We identified candidate promoters from Arabidopsis based on their transcript levels (meta-analysis of avail...
journal_title:Transgenic research
pub_type: 杂志文章,meta分析
doi:10.1007/s11248-013-9757-9
更新日期:2014-04-01 00:00:00
abstract::The disease model of familial amyloidotic polyneuropathy-7.2-hMet30 mice-manifests amyloid deposition that consists of a human amyloidogenic mutant transthyretin (TTR) (TTR V30M). Our previous study found amyloid deposits in 14 of 27 7.2-hMet30 mice at 21-24 months of age. In addition, non-fibrillar TTR deposits were ...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-017-0054-x
更新日期:2018-02-01 00:00:00
abstract::To construct an efficient system for the production of recombinant proteins in silkworm (Bombyx mori), we investigated the promoter activity of the silkworm sericin 1, 2, and 3 genes (Ser1, Ser2, and Ser3) using a GAL4/UAS binary gene expression system in transgenic silkworm. The promoter activity of the upstream regi...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-009-9328-2
更新日期:2010-06-01 00:00:00
abstract::To identify ways to improve the efficiency of generating chimeric mice via microinjection of blastocysts with ES cells, we compared production and performance of ES-cell derived chimeric mice using blastocysts from two closely related and commonly used sub-strains of C57BL/6. Chimeras were produced by injection of the...
journal_title:Transgenic research
pub_type: 信件
doi:10.1007/s11248-012-9605-3
更新日期:2012-12-01 00:00:00
abstract::Green fluorescent protein (GFP) transgenic animals are widely used in biomedical research. We observed that the commonly used beta-actin-GFP transgenic mouse has renal defects with proteinuria starting as early as 5 weeks of age. Histological analysis reveals a widespread increase in glomerular extracellular matrix, o...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-007-9107-x
更新日期:2007-12-01 00:00:00
abstract::Short directly-repeated (DR) DNA enhancer elements of plant viral origin were analyzed for their ability, both individually and in combination, to influence in vivo transcription when inserted upstream from a minimal CaMV35S promoter. Synthetic promoters containing multiple copies and/or combinations of DR cassettes w...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/s11248-007-9117-8
更新日期:2008-06-01 00:00:00
abstract::In the fowl the primordial germ cells accumulate in the germinal crescent to the anterior of the two-day embryo. A simple ballistic device has been used to fire tungsten particles (mean diameter 1.5 microns) into this region. By coating these projectiles with vector DNA it is possible to transfect these cells. Hatchli...
journal_title:Transgenic research
pub_type: 杂志文章
doi:10.1007/BF01976498
更新日期:1995-01-01 00:00:00