specific gene silencing using rnai in cell culture

... Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site -specific Cre recombinase Nucleic Acids Res 1995, 23(19):3816-3821 Moffat JF, Zerboni L, Kinchington PR, ... suggesting that the ORF58 gene is dispensable for viral replication in cell culture Before performing the remaining experiments, in order to exclude the possibility to affect the packaging of ... was amplified from the rpOka-infected cells, but not from the pOka∆58-infected cells, indicating that the ORF58 gene product was not expressed in the rpOka∆58-infected cells As positive controls,...

Ngày tải lên: 20/06/2014, 01:20

... budding Binding Protein regions presumably CBD aa47-55 Binding to CSD*† Binding to CSD‡ This investigation Zou et al 2009 Influenza A virus human HA Receptor binding Colocalization in perinuclear ... caveolin-1 binding domain in human influenza A viruses A For reasons of clearness, only a M2 monomer is indicated in the drawing M2 tetramers function as an ion-pump (residing in a helical domain in ... represented by cylinder 1) The C-terminal region is important for virus assembly and budding A palmitoyl residue (jigsaw line) is linked to cysteine 50 The caveolin-1 binding domain resides in the loop...

Ngày tải lên: 12/08/2014, 04:20

... in viral inactivation Ten minutes of glutaraldehyde treatment resulted in more than 1-log reduction in infectivity After 20 min, the virus stock was inactivated completely, with no residual infectivity ... results in FFU assay or at the second cell passage were not tested for NS3 staining for the third cell passage from any of these cell lysates in three consecutive passages of inoculated cells (Table ... HCVcc inactivation took place immediately after vortex-mixing, rendering it impossible to delineate the kinetics of viral infectivity reduction during the detergent treatment process This finding...

Ngày tải lên: 12/08/2014, 04:21

... 0.23 in Generation to 0.57 in Generation in the absence of thyroid hormone and from 0.05 in Generation to 0.11 in Generation in its presence Therefore, the survival rate of O-2A/OPCs increases in ... probability of differentiation increases from 0.07 in Generation to 0.21 in Generation in the absence of thyroid hormone and from 0.18 in Generation to 0.72 in Generation in its presence This is consistent ... basic assumptions behind our model of oligodendrocyte development in cell culture In what follows, we describe our experimental findings in the context of the model presented in Section Each element...

Ngày tải lên: 13/08/2014, 23:20

... and CyclinD1 had no distinct difference in three groups(P > 0.05) (Figure 4) MTA1 silencing reduces the invasive ability of MDA-MB231 cells in vitro The effects of inhibiting MTA1 gene on invasion ... After silencing of MTA1 gene, we investigated the alteration of tumor cells invasiveness using Boyden chamber assay mentioned in Albini’s[20] literature The results showed that tumor cells invasiveness ... transcribe and generate siRNA continuously and the gene silencing effect can last persistently inside the cells These findings have opened a broad new avenue for the analysis of gene function and gene...

Ngày tải lên: 10/08/2014, 10:21

... These data indicated that JMJD2A -specific siRNA silencing mRNA could significantly reduce the levels of JMJD2A protein expression in MDA-MB-231 cells Silencing JMJD2A gene resulted in cell cycle ... expressed in human tissues and cell lines, and high endogenous expression of JMJD2A mRNA was found in several cell types, including human T -cell lymphotropic virus 1infected cell lines, the HT1376 ... cotton swab The cells that migrated to the bottom surface of the insert were fixed in 100% methanol for min, stained in Giemsa for min, rinsed in PBS and then subjected to microscopic inspection (×200)...

Ngày tải lên: 10/08/2014, 10:21

... including establishing, selecting, and maintaining all transduced cell lines AK and RD were instrumental in providing support to the research in the form of both instruction in and preparation of all ... transduced cells were maintained in complete L-15 with 35 μg/mL hygromycin (Invitrogen) For infection of cells, complete L-15 without any antibiotics was used Cells were incubated at 28°C without CO2 Infection ... hRz expressing C6/36 cells using qRT-PCR to detect virus genomes in cell lysates Total cellular RNA was extracted dpi from infected hRz-transduced cells following the protocol described in Materials...

Ngày tải lên: 12/08/2014, 04:21

... of 10 Figure Changes in gene expression during passaging (a) The changes in gene expression increase substantially during passaging of the cells in all fibroblast cultures In addition, the variations ... approved the final manuscript Figure Cell doubling rates (in days) during cell culture passages for each patient The cell doubling rate was measured by counting the cells The day, the cell numbers ... starting in a defined passage reaches statistical significance and differential expression continues through passaging of the cell population (>'passage') In addition, two genes were verified using...

Ngày tải lên: 12/08/2014, 12:20

... 1: Introduction Chapter Introduction Chapter 1: Introduction Introduction 1.1 Overview of Gene Therapy Somatic gene therapy is based upon the introduction of therapeutically active genes into individual ... results in large increase in the proton concentration 12 Chapter 1: Introduction within the endosome, resulting in swelling of the polymer by internal charge repulsion, with resultant osmotic swelling ... condense the DNA further with increased packing of amines around the DNA 2.2 Cell Targeting Using Nerve Growth Factor For many envisioned applications in gene delivery, a gene delivery vehicle should...

Ngày tải lên: 27/11/2015, 11:11

... Berns A (2002) Tumor formation in mice with somatic inactivation of the retinoblastoma gene in interphotoreceptor retinal binding protein-expressing cells Oncogene 21:4635–4645 Voorhoeve PM, ... 14:2358–2365 Weinberg RA (1995) The retinoblastoma protein and cell cycle control Cell 81:323–330 Welch PJ, Wang JY (1993) A C-terminal protein-binding domain in the retinoblastoma protein regulates ... while increased apoptosis was only observed in follicular cells lacking both pRb and p107 (Ruiz et al 2004) Interesting observations were made in cells lacking pRb in the retina Somatic inactivation...

Ngày tải lên: 25/10/2013, 21:20

... above) Stock cell culture and culture condition Suspended cells of P vietnamensis were initiated through callus induction from the cultivated plant root [5] The cell line was maintained in MS liquid ... of ginseng cell and some attempts have been made to increase biomass and ginsenoside yield of Ngoc Linh ginseng cell culture Materials and methods Induction of callus Fresh mountain ginseng roots ... In cell cultures of P quinquefolium, [12] reported that the final dry cell weight was relatively low with the low nitrogen concentration Maximum cell dry weight obtained (15 g/L) at a total initial...

Ngày tải lên: 12/02/2014, 10:20

... et al Interspecies comparison of ecdysteroid receptors Fig EMSA using CHO cell extracts following transfection and incubation in the absence and presence of MurA, RH5849, and RH5992, using the ... capable of mimicking or evoking the potentiation effect induced by JHIII in this assay Experimental procedures Cell culture, EMSA, and western immunoblotting All aspects of cell culture methodology, ... sites inserted by the PCR primers are underlined The resulting LdUSP amplicon includes the entire DBD, whereas LdUSP ⁄ DDBD includes the entire ORF beginning at the first amino acid following the...

Ngày tải lên: 18/02/2014, 08:20

... nutrition and insulin signaling is inverted in honey bee adults, and JH inhibits Vg expression in adults rather than stimulating it [59,60] The short neuropeptide F signaling cascade is involved in fire ... Streptavidin (Invitrogen; : 200) in blocking solution for h Sections were washed and mounted in Vectashield Mounting medium with 4¢,6-diamidino-2-phenylindole for nuclear staining (Vector, Burlingame, ... higher in the ovary of the egg-laying queen within a mature colony than in virgin queens [12] The subcellular localization of SiVgR signal was similar in virgin and mated queens, i.e expressed in...

Ngày tải lên: 18/02/2014, 08:20

... N-terminus of the GUS gene and inserted into pTF1SIIn to replace the NtFAD3 sequence Then, a cassette containing a nopaline synthase (Nos) promoter and hygromycin phosphotransferase B gene was inserted ... gene silencing Plant Mol Biol 43, 243–260 11 Ma C & Mitra A (2002) Intrinsic direct repeats generate consistent post-transcriptional gene silencing in tobacco Plant J 31, 37–49 12 Tang G, Reinhart ... regulation in S-PTGS S Hirai et al is restored in C002 plants [30,31] Similarly, S-PTGS of the neomycin phosphotransferase II gene (nptII) is eliminated during regeneration from in vitro-cultured cells...

Ngày tải lên: 22/03/2014, 21:20

... specifically in CA until the beginning of the final larval instar [9] In contrast, the BmJHAMT transcript is undetectable in CA at the beginning of the spinning stage, but is detected at low levels in the ... recombinant TcMT3 protein (B) Arrows indicate (10S)-JH III and (10R)-JH III TcMT1 dsRNA was injected in the prepupal stage and TcMT2 dsRNA was injected at the beginning of the final larval instar In ... methyltransferase genes in the larval stage in vivo, RNAi- mediated knockdown of TcMT1, TcMT2 and TcMT3 was performed by injecting dsRNA at the beginning of the third instar A B C D Fig In situ hybridization...

Ngày tải lên: 23/03/2014, 07:20

... [1] reported that NH4+ in the culture medium inhibited ginsenoside accumulation in P notoginseng cell suspension cultures and that maximum ginsenoside production was obtained when NH4+ was absent ... cell culture and culture condition Suspended cells of P vietnamensis were initiated through callus induction from the cultivated plant root [8] The cell line was maintained in MS liquid medium ... obtained when 0.5 strength level of NH4NO3 from the culture medium Table Biomass growth of ginseng cell was affected by concentration of macro elements in the MS medium Cultures were maintained in...

Ngày tải lên: 28/03/2014, 10:20

... sample of cell lines using DNA fingerprinting and cytogenetic evaluation have shown a high incidence (approx 15%) of false cell lines observed among cell lines obtained directly from original investigators ... enabling every researcher using human cell lines to easily verify cell line identity Key Words: Authentication; cross-contamination; PCR; DNA fingerprinting; false cell lines; VNTR; AmpFLP Introduction ... Basic Cell Culture Techniques 1 Culture of Primary Adherent Cells and a Continuously Growing Nonadherent Cell Line Cheryl D Helgason Summary Cell culture is an invaluable tool for investigators in...

Ngày tải lên: 08/04/2014, 12:50

... resuspended in 50 μl of staining solution (PBS, 3% FBS and 0,1% NaN3) containing an optimal concentration of antibody After a 20-minute incubation in ice, cells were washed again and analyzed using a ... values from independent experiments carried out for each donor cell line of cells required (as few as 0.32 × 10 cells for each test), in this case we could test every cell line obtained In line with ... (Figure 4b) These findings are in line with our previous observations [13] and the vast majority of data related to EBV -specific cultures [20] Once again, cytokines used in cultures did not modify...

Ngày tải lên: 18/06/2014, 16:20

... bases in the minus-sense RNA Interestingly, in Puumala hantavirus, a hairpin-like structure formed by a highly conserved inverted repeat in the 3'-noncoding region of the S segment seems to be involved ... RNA from cells infected with TULV02 (line 1), on RNA from cells infected with the recTULV (line 2) and on the mechanical mixture of both RNA preparations (line 3) Lines 5–7: the corresponding products ... O: Intragenic recombination in rotaviruses FEBS Letters 1998, 427:183-187 Nayak DP, Chambers TM, Akkina RK: Defective interfering RNAs of influenza viruses: origin, structure, expression and interference...

Ngày tải lên: 18/06/2014, 22:20