Detection of Antisense RNA Transcripts by Strand-Specific RT-PCR

2010 ◽  
pp. 125-138 ◽  
Author(s):  
Eric C. H. Ho ◽  
Michael E. Donaldson ◽  
Barry J. Saville
Keyword(s):  
Antisense Rna ◽  
Rt Pcr ◽  
Rna Transcripts

scholarly journals Detection Method for Multiple Gene Expression in Single Early Bovine Embryo (Antisense RNA-RT-PCR).

1996 ◽  
Vol 13 (2) ◽  
pp. 125-129
Author(s):  
Hiroya Kadokawa ◽  
Seizo Hamano ◽  
Ryuji Itoh ◽  
Hitomi Takahashi ◽  
Yutaka Yamada ◽  
...  
Keyword(s):  
Gene Expression ◽  
Antisense Rna ◽  
Detection Method ◽  
Bovine Embryo ◽  
Rt Pcr ◽  
Multiple Gene

Microinjection of fos-specific antibodies blocks DNA synthesis in fibroblast cells

1988 ◽  
Vol 8 (4) ◽  
pp. 1670-1676 ◽  
Author(s):  
K T Riabowol ◽  
R J Vosatka ◽  
E B Ziff ◽  
N J Lamb ◽  
J R Feramisco
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Antisense Rna ◽  
Proliferating Cells ◽  
Rna Transcripts ◽  
Serum Stimulation ◽  
Inhibition Of Growth ◽  
Nonspecific Immunoglobulins ◽  
3T3 Cell Lines ◽  
Stimulation Of

Transcription of the protooncogene c-fos is increased greater than 10-fold within minutes of treatment of fibroblasts with serum or purified growth factors. Recent experiments with mouse 3T3 cell lines containing inducible fos antisense RNA constructs have shown that induced fos antisense RNA transcripts cause either a marked inhibition of growth in continuously proliferating cells or, conversely, a minimal effect except during the transition from a quiescent (G0) state into the cell cycle. Since intracellular production of large amounts of antisense RNA does not completely block gene expression, we microinjected affinity-purified antibodies raised against fos to determine whether and when during the cell cycle c-fos expression was required for cell proliferation. Using this independent method, we found that microinjected fos antibodies efficiently blocked serum-stimulated DNA synthesis when injected up to 6 to 8 h after serum stimulation of quiescent REF-52 fibroblasts. Furthermore, when fos antibodies were injected into asynchronously growing cells, a consistently greater number of cells was prevented from synthesizing DNA than when cells were injected with nonspecific immunoglobulins. Thus, whereas the activity of c-fos may be necessary for transition of fibroblasts from G0 to G1 of the cell cycle, its function is also required during the early G1 portion of the cell cycle to allow subsequent DNA synthesis.

scholarly journals Translation of mRNA injected into Xenopus oocytes is specifically inhibited by antisense RNA.

1985 ◽  
Vol 101 (3) ◽  
pp. 1094-1099 ◽  
Author(s):  
R Harland ◽  
H Weintraub
Keyword(s):  
Rna Polymerase ◽  
Antisense Rna ◽  
Xenopus Oocytes ◽  
Oocyte Nucleus ◽  
Detectable Effect ◽  
Antisense Rnas ◽  
Rna Transcripts ◽  
Oocyte Cytoplasm ◽  
Endogenous Proteins ◽  
Specific Mrna

The bacteriophage SP6 promoter and RNA polymerase were used to synthesize sense and antisense RNAs coding for the enzymes thymidine kinase (TK) and chloramphenicol acetyl transferase (CAT). Injection of antisense CAT RNA into frog oocytes inhibited expression of sense CAT mRNA. Similarly, antisense TK RNA inhibited expression of sense TK mRNA. Antisense RNAs were stable in oocytes and had no detectable effect on either the expression of endogenous proteins or on the expression of nonhomologous RNA transcripts. CAT activity expressed from a plasmid transcribed in the oocyte nucleus was also inhibited by antisense RNA injected into the oocyte cytoplasm. The data suggest that antisense RNA will be useful in identifying the function of specific mRNA sequences during early development of the frog.

scholarly journals Direct Quantitation of RNA Transcripts by Competitive Single-Tube RT-PCR and Capillary Electrophoresis

BioTechniques ◽  
1998 ◽  
Vol 25 (1) ◽  
pp. 130-137 ◽  
Author(s):  
N.D. Borson ◽  
M.A. Strausbauch ◽  
P.J. Wettstein ◽  
R.P. Oda ◽  
S.L. Johnston ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Rt Pcr ◽  
Single Tube ◽  
Rna Transcripts

scholarly journals Presence and distribution of human papillomavirus sense and antisense RNA transcripts in genital cancers

1991 ◽  
Vol 72 (4) ◽  
pp. 885-895 ◽  
Author(s):  
G. D. Higgins ◽  
D. M. Uzelin ◽  
G. E. Phillips ◽  
C. J. Burrell
Keyword(s):  
Human Papillomavirus ◽  
Antisense Rna ◽  
Rna Transcripts

Inhibition of Potato Spindle Tuber Viroid (PSTVd) Infectivity with Antisense RNA Transcripts

1994 ◽  
Vol 141 (1) ◽  
pp. 10-24 ◽  
Author(s):  
J. Matoušek ◽  
L. Trněná ◽  
S. Rakouský ◽  
D. Riesner
Keyword(s):  
Antisense Rna ◽  
Potato Spindle Tuber Viroid ◽  
Rna Transcripts

scholarly journals Origin, Splicing, and Expression of Rodent Amelogenin Exon 8

2006 ◽  
Vol 85 (10) ◽  
pp. 894-899 ◽  
Author(s):  
J.D. Bartlett ◽  
R. L. Ball ◽  
T. Kawai ◽  
C.E. Tye ◽  
M. Tsuchiya ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Splice Site ◽  
Site Selection ◽  
Proteolytic Processing ◽  
Rt Pcr ◽  
Splice Site Selection ◽  
Rna Transcripts ◽  
Splicing Pattern ◽  
Alternatively Spliced ◽  
Gene Structures

Amelogenin RNA transcripts undergo extensive alternative splicing, and MMP-20 processes the isoforms following their secretion. Since amelogenins have been ascribed cell-signaling activities, we asked if a lack of proteolytic processing by MMP-20 affects amelogenin signaling and consequently alters amelogenin splice site selection. RT-PCR analyses of amelogenin mRNA between control and Mmp20− /−mice revealed no differences in the splicing pattern. We characterized 3 previously unidentified amelogenin alternatively spliced transcripts and demonstrated that exon-8-encoded amelogenin isoforms are processed by MMP-20. Transcripts with exon 8 were expressed approximately five-fold less than those with exon 7. Analyses of the mouse and rat amelogenin gene structures confirmed that exon 8 arose in a duplication of exons 4 through 5, with translocation of the copy downstream of exon 7. No downstream genomic sequences homologous to exons 4–5 were present in the bovine or human amelogenin genes, suggesting that this translocation occurred only in rodents.

scholarly journals Detection of Metalloproteases and Cysteine Proteases RNA Transcripts of Leishmania (Leishmania) infantum in Ear Edge Skin of Naturally Infected Dogs

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Laura Barral Veloso ◽  
Flávia de Oliveira Cardoso ◽  
Karen dos Santos Charret ◽  
Matheus Pereira de Sá Silva ◽  
Luzia Monteiro de Castro Côrtes ◽  
...  
Keyword(s):  
Gene Expression ◽  
Virulence Factors ◽  
Leishmania Infantum ◽  
Natural Infection ◽  
Parasite Load ◽  
Cysteine Proteases ◽  
Rt Pcr ◽  
Load Range ◽  
Rna Transcripts ◽  
Pcr Assays

Leishmania spp. proteases have been proposed as virulence factors contributing to adaptive success these parasites to the mammalian hosts. Since these enzymes are poorly studied in naturally infected dogs, this work aims to show the differences in metalloprotease and cysteine proteases gene expression in ear edge skin of dogs naturally infected by Leishmania (Leishmania) infantum. A cohort of dogs (n=20) naturally infected by L. (L.) infantum was clinically classified as asymptomatic, oligosymptomatic, and polysymptomatic and the parasite load range estimated. The analysis of proteases expression by RT-PCR in the ear edge skin was also assessed, suggesting more transcripts of proteases in cDNA samples from polysymptomatic dogs than oligosymptomatic and asymptomatic ones. Metalloprotease RT-PCR assays yielded products (202 bp) in all assessed cDNA dog samples. In contrast, cysteine proteases transcripts (227 bp) had shown to be better detected in cDNA samples of polysymptomatic dogs, compared with cDNA samples from asymptomatic and oligosymptomatic dogs. Predictive in silico assays suggested that secondary structures of metalloproteasee mRNAs can be more stable than cysteine proteases at the skin temperature of dogs. Evidence is presented that during natural infection of dogs by L. (L.) infantum, this parasite produces transcripts of metalloprotease and cysteine protease RNA in the skin from asymptomatic, oligosymptomatic, and polysymptomatic dogs.

scholarly journals ADAR2-Mediated Editing of miR-214 and miR-122 Precursor and Antisense RNA Transcripts in Liver Cancers

PLoS ONE ◽  
2013 ◽  
Vol 8 (12) ◽  
pp. e81922 ◽  
Author(s):  
Wan-Hsin Liu ◽  
Chao-Hung Chen ◽  
Kun-Huei Yeh ◽  
Chiao-Ling Li ◽  
Yi-Jinn Wu ◽  
...  
Keyword(s):  
Antisense Rna ◽  
Rna Transcripts ◽  
Liver Cancers
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