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Transient assay
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The CRISPR/Cas9 system has recently been engineered to confer resistance to geminiviruses in plants. However, we show here that the usefulness of this antiviral strategy is undermined by off-target effects identified by deep sequencing in Arabidopsis. We construct two virus-inducible CRISPR/Cas9 vectors that efficiently inhibit beet severe curly top virus (BSCTV) accumulation in both transient assays (Nicotiana benthamiana) and transgenic lines (Arabidopsis).
7p
vigalileogalilei
27-02-2022
28
1
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Soybean homeobox gene GmSBH1 has previously been proven to be involved in response to high temperature and humidity (HTH) stress. To investigate its expression patterns and active cis-elements, a 2040-bp 5’-upstream genomic DNA fragment of GmSBH1, named GmSBH1P, was isolated by PCR walking and characterized. Sequence analysis revealed that the fragment contains a series of cis-acting elements related to stress responses.
13p
tudichquannguyet
29-11-2021
9
1
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The combination of time-lapse imaging of live cells with high-throughput perturbation assays is a powerful tool for genetics and cell biology. The Mitocheck project employed this technique to associate thousands of genes with transient biological phenotypes in cell division, cell death and migration.
10p
viwyoming2711
16-12-2020
8
1
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A simple method for transient expression of reporter gene in Brinjal leaves through agroinfiltration
Brinjal (Solanum melongena L.) is one of the most important vegetable crops in India. In spite of the docility of brinjal towards genetic transformation through Agrobacterium, development of stable transgenic lines in brinjal is a cost, labour as well as time consuming process. On the other hand, the agro infiltration process has been optimized in different crops for the rapid analysis of transgene through transient expression. The process has also gained importance in synthetic biological applications for rapid production of therapeutic biomolecules in plants.
7p
chauchaungayxua6
26-06-2020
24
1
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Transcriptional enhancers are able to increase transcription from heterologous promoters when placed upstream, downstream and in either orientation, relative to the promoter.
12p
viminato2711
22-05-2020
11
1
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Agrobacterium tumefaciens-based transient assays have become a common tool for answering questions related to protein localization and gene expression in a cellular context. The use of these assays assumes that the transiently transformed cells are observed under relatively authentic physiological conditions and maintain ‘normal’ sub-cellular behaviour.
20p
viminato2711
22-05-2020
10
1
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In this study we describe the isolation of three genes of the CYP11B family of the guinea pig. CYP11B1 codes for the previously described 11b-hydroxylase [Bulow, H.E., Mobius, ¨ ¨ K., Bahr, V. & Bernhardt, R. (1996) Biochem. Biophys. Res. ¨ Commun. 221, 304–312] while CYP11B2 represents the aldosterone synthase gene. As no expression for CYP11B3 was detected this gene might represent a pseudogene.
9p
system191
01-06-2013
36
2
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The CD53 antigen is a member of the tetraspanin membrane protein family that is expressed in the lymphoid-myeloid lineage. We have studied the implication of CD53 antigen in signal transduction by determining the effect of its ligation on the c-Jun N-terminal kinase (JNK) in different cell types. Ligation of the rat or human CD53 antigen induces a threeto fourfold transient activation of JNK activity that peaks at 3–5 min. The effect was detected by assaying the endogenous or exogenous (transfected) JNK activity....
10p
research12
01-06-2013
38
4
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Glutamine synthetase (GS) is expressed at high levels in subsets of cells in some tissues and at low levels in all cells of other tissues, suggesting that the GS gene is surrounded by multiple regulatory elements.We searched for such elements in the 2.5-kbupstreamregionand in the 2.6-kbfirst intronof theGSgene, usingFTO-2Bhepatoma andC2/7muscle cells as representatives of both cell types and transient transfec-tion assays as our tools.
7p
tumor12
20-04-2013
34
2
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Mitochondrial biogenesis is a complex and highly regulated process that requires the controlled expression of hundreds of genes encoded in two separated genomes, namely the nuclear and mitochondrial genomes. To identify regulatory proteins involved in the transcriptional control of key nuc-lear-encoded mitochondrial genes, we have performed a detailed analysis of the promoter region of theasubunit of the Drosophila melanogasterF1F0 ATP synthase complex.
11p
awards
05-04-2013
37
3
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To identify artificial DNA segments that can stably express transgenes in the genome of host cells, we built a series of curved DNA segments that mimic a left-handed superhelical structure. Curved DNA segments of 288 bp (T32) and 180 bp (T20) were able to activate transcription from the herpes simplex virus thymidine kinase (tk) promoter by approximately 150-fold and 70-fold, respectively, compared to a control in a transient transfection assay in COS-7 cells.
12p
inspiron33
23-03-2013
41
2
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