Crosslinking and immunoprecipitation (CLIP) strategies are highly effective strategies to interrogate direct protein-RNA interactions and dissect posttranscriptional gene regulatory networks. One broadly used CLIP variant is photoactivatable ribonucleoside enhanced CLIP (PAR-CLIP) that entails in vivo labeling of nascent RNAs with the photoreactive nucleosides 4-thiouridine (4SU) or 6-thioguanosine (6SG), which might effectively crosslink to interacting proteins utilizing UVA and UVB mild.
Crosslinking of 4SU or 6SG to interacting amino acids adjustments their base-pairing properties and leads to attribute mutations in cDNA libraries ready for high-throughput sequencing, which could be computationally exploited to take away ample background from non-crosslinked sequences and assist pinpoint RNA binding protein binding websites at nucleotide decision on a transcriptome-wide scale.
Right here we current a streamlined protocol for fluorescence-based PAR-CLIP (fPAR-CLIP) that eliminates the necessity to use radioactivity. It’s primarily based on direct ligation of a fluorescently labeled adapter to the three’finish of crosslinked RNA on immobilized ribonucleoproteins, adopted by isolation of the adapter-ligated RNA and environment friendly conversion into cDNA with out the beforehand wanted measurement fractionation on denaturing polyacrylamide gels. These enhancements reduce the experimentation by half to 2 days and will increase sensitivity by 10-100-fold.
Characterization and evaluation of the transcriptome response to drought in Larix kaempferi utilizing PacBio full-length cDNA sequencing built-in with de novo RNA-seq reads
A hypothetical mannequin of drought tolerance mechanism of Larix kaempferi was established via SMRT-seq and Illumina HiSeq. Larix kaempferi is a vital financial and ecological species and a significant afforestation species in north-eastern China. Up to now, no info has been reliably derived concerning full-length cDNA sequencing info on L. kaempferi. By single-molecule long-read isoform sequencing (SMRT-seq), right here we report a complete of 26,153,342 subreads (21.24 Gb) and 330,371 round consensus sequence (CCS) reads after the modification of web site mismatch, and 35,414 unigenes have been efficiently collected.
To achieve deeper insights into the molecular mechanisms of L. kaempferi response to drought stress, we mixed Illumina HiSeq with SMRT-seq to decode full-length transcripts. On this examine, we report 27 differentially expressed genes (DEGs) concerned within the notion and transmission of drought stress indicators in L. kaempferi.
Numerous DEGs responding to drought stress have been detected in L. kaempferi, particularly DEGs concerned within the reactive oxygen species (ROS) scavenging, lignin biosynthesis, and sugar metabolism, and DEGs encoding drought stress proteins. We detected 73 transcription components (TFs) below drought stress, together with AP2/ERF, bZIP, TCP, and MYB. This examine offers primary full sequence sources for L. kaempferi analysis and can assist us to raised perceive the capabilities of drought-resistance genes in L. kaempferi.
cDNA-derived RNA phage meeting reveals important residues within the maturation protein of the Pseudomonas aeruginosa leviphage, PP7
PP7 is a leviphage with single-stranded RNA genome, which infects Pseudomonas aeruginosa PAO1. A reverse genetic system for PP7 was beforehand created through the use of reverse-transcribed cDNA (PP7O) from virion-derived RNA genome. Right here, we’ve discovered that the PP7O cDNA contained 20 nucleotide variations from the PP7 genome sequence deposited within the database.
We created one other reverse genetic system exploiting chemically synthesized cDNA (PP7S) primarily based on the database sequence. In contrast to PP7O that rendered infectious PP7 virions, PP7S-derived particles have been incapable of plaque formation on PAO1 cells, which was restored on the PAO1 cells expressing the maturation protein (MP) from PP7O Utilizing this reverse genetic system, we revealed two amino acid residues concerned within the identified roles of MP (i.e. adsorption and genome replication), fortuitously offering a lesson that the viral RNA genome sequencing wants purposeful verification presumably by a reverse genetic system.
IMPORTANCE Organic significance of RNA phages has been largely ignored, mockingly as a result of few research have been specializing in RNA phages. As an preliminary try to correctly signify RNA phages within the phageome, we beforehand created, through the use of reverse-transcribed cDNA, a reverse genetic system for the small RNA phage, PP7 that infects the opportunistic human pathogen, Pseudomonas aeruginosa We right here report one other system through the use of chemically synthesized cDNA primarily based on the database genome that has 20 nucleotide variations from the earlier cDNA.
Investigation of these cDNA-derived phage virions unveiled that two amino acids of the maturation protein are essential for the traditional phage lifecycle at totally different steps. Our examine offers an perception into the molecular foundation for the RNA phage lifecycle and a lesson that the RNA genome sequencing must be rigorously validated by cDNA-based phage meeting methods.
RNA–cDNA hybrids mediate transposition by way of totally different mechanisms
Retrotransposons can signify half of eukaryotic genomes. Retrotransposon dysregulation destabilizes genomes and has been linked to varied human ailments. Rising regulators of retromobility embrace RNA-DNA hybrid-containing constructions generally known as R-loops. Accumulation of those constructions on the transposons of yeast 1 (Ty1) parts has been proven to extend Ty1 retromobility via an unknown mechanism. Right here, by way of a focused genetic display screen, we recognized the rnh1Δ rad27Δ yeast mutant, which lacked each the Ty1 inhibitor Rad27 and the RNA-DNA hybrid suppressor Rnh1.
The mutant exhibited elevated ranges of Ty1 cDNA-associated RNA-DNA hybrids that promoted Ty1 mobility. Furthermore, on this rnh1Δ rad27Δ mutant, however not within the double RNase H mutant rnh1Δ rnh201Δ, RNA-DNA hybrids preferentially existed as duplex nucleic acid constructions and elevated Ty1 mobility in a Rad52-dependent method.
 POLR3F siRNA |
|
20-abx929231 |
Abbexa |
|
|
|
|
POLR3F siRNA |
|
20-abx929232 |
Abbexa |
|
|
|
|
 POLR3F Peptide |
|
4727P |
ProSci |
0.05 mg |
EUR 197.7 |
|
Description: (NT) POLR3F peptide |
 POLR3F Antibody |
|
24726-100ul |
SAB |
100ul |
EUR 468 |
 POLR3F antibody |
|
70R-2962 |
Fitzgerald |
50 ug |
EUR 560.4 |
|
Description: Rabbit polyclonal POLR3F antibody raised against the N terminal of POLR3F |
POLR3F antibody |
|
70R-19415 |
Fitzgerald |
50 ul |
EUR 522 |
|
Description: Rabbit polyclonal POLR3F antibody |
POLR3F antibody |
|
70R-2038 |
Fitzgerald |
50 ug |
EUR 560.4 |
|
Description: Rabbit polyclonal POLR3F antibody raised against the middle region of POLR3F |
POLR3F antibody |
|
70R-50816 |
Fitzgerald |
100 ul |
EUR 292.8 |
|
Description: Purified Polyclonal POLR3F antibody |
POLR3F Antibody |
|
4727-002mg |
ProSci |
0.02 mg |
EUR 206.18 |
|
|
|
Description: POLR3F Antibody: The human POLR3F is a component of RNA III polymerase. RNA polymerase III transcribes many essential, small, noncoding RNAs, including the 5S rRNAs and tRNAs. While most pol III-transcribed genes are found scattered throughout the linear chromosome maps or in multiple linear clusters, there is increasing evidence that many of these genes prefer to be spatially clustered, often at or near the nucleolus. This association could create an environment that fosters the coregulation of transcription by pol III with transcription of the large ribosomal RNA repeats by RNA polymerase I (pol I) within the nucleolus. Given the high number of pol III-transcribed genes in all eukaryotic genomes, the spatial organization of these genes is likely to affect a large portion of the other genes in a genome. POLR3F has also been recently identified as an HIV dependency factor (HDF), suggesting that POLR3F may be an important drug target in HIV treatment. At least two isoforms of POLR3F are known to exist. |
POLR3F Antibody |
|
4727-01mg |
ProSci |
0.1 mg |
EUR 523.7 |
|
|
|
Description: POLR3F Antibody: The human POLR3F is a component of RNA III polymerase. RNA polymerase III transcribes many essential, small, noncoding RNAs, including the 5S rRNAs and tRNAs. While most pol III-transcribed genes are found scattered throughout the linear chromosome maps or in multiple linear clusters, there is increasing evidence that many of these genes prefer to be spatially clustered, often at or near the nucleolus. This association could create an environment that fosters the coregulation of transcription by pol III with transcription of the large ribosomal RNA repeats by RNA polymerase I (pol I) within the nucleolus. Given the high number of pol III-transcribed genes in all eukaryotic genomes, the spatial organization of these genes is likely to affect a large portion of the other genes in a genome. POLR3F has also been recently identified as an HIV dependency factor (HDF), suggesting that POLR3F may be an important drug target in HIV treatment. At least two isoforms of POLR3F are known to exist. |
POLR3F Antibody |
|
1-CSB-PA884440LA01HU |
Cusabio |
|
|
|
|
|
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is Unconjugated. Tested in the following application: ELISA, WB; Recommended dilution: WB:1:500-1:2000 |
POLR3F Antibody |
|
1-CSB-PA018348GA01HU |
Cusabio |
|
|
|
|
|
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB |
POLR3F Antibody |
|
E94441 |
EnoGene |
100μg |
EUR 255 |
|
Description: Available in various conjugation types. |
POLR3F Antibody |
|
E38PA2497 |
EnoGene |
100ul |
EUR 225 |
|
Description: Available in various conjugation types. |
POLR3F Antibody |
|
E912224 |
EnoGene |
100ul |
EUR 255 |
|
Description: Available in various conjugation types. |
 POLR3F Rabbit pAb |
|
A12224-100ul |
Abclonal |
100 ul |
EUR 369.6 |
POLR3F Rabbit pAb |
|
A12224-200ul |
Abclonal |
200 ul |
EUR 550.8 |
POLR3F Rabbit pAb |
|
A12224-20ul |
Abclonal |
20 ul |
EUR 219.6 |
POLR3F Rabbit pAb |
|
A12224-50ul |
Abclonal |
50 ul |
EUR 267.6 |
POLR3F Rabbit pAb |
|
E2510867 |
EnoGene |
100ul |
EUR 225 |
|
Description: Available in various conjugation types. |
 anti- POLR3F antibody |
|
FNab06632 |
FN Test |
100µg |
EUR 658.5 |
|
|
|
Description: Antibody raised against POLR3F |
 POLR3F Blocking Peptide |
|
20-abx063691 |
Abbexa |
|
|
|
|
POLR3F Blocking Peptide |
|
33R-5238 |
Fitzgerald |
100 ug |
EUR 216 |
|
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of POLR3F antibody, catalog no. 70R-2038 |
POLR3F Blocking Peptide |
|
33R-5647 |
Fitzgerald |
100 ug |
EUR 216 |
|
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of POLR3F antibody, catalog no. 70R-2962 |
 POLR3F Polyclonal Antibody |
|
27655-100ul |
SAB |
100ul |
EUR 302.4 |
POLR3F Polyclonal Antibody |
|
27655-50ul |
SAB |
50ul |
EUR 224.4 |
POLR3F Polyclonal Antibody |
|
A60346 |
EpiGentek |
-
EUR 684.66
-
EUR 117.70
-
EUR 302.50
-
EUR 423.50
|
- 100 µg
- 20 ul
- 50 ul
- 100 ul
|
) POLR3F protein (His tag) |
|
30R-2961 |
Fitzgerald |
100 ug |
EUR 386.4 |
|
Description: Purified recombinant Human POLR3F protein (His tag) |
 Human POLR3F shRNA Plasmid |
|
20-abx957233 |
Abbexa |
|
|
|
|
 Mouse POLR3F shRNA Plasmid |
|
20-abx977203 |
Abbexa |
|
|
|
|
 POLR3F Antibody, HRP conjugated |
|
1-CSB-PA884440LB01HU |
Cusabio |
|
|
|
|
|
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is HRP conjugated. Tested in the following application: ELISA |
) POLR3F Recombinant Protein (Rat) |
|
RP221387 |
ABM |
100 ug |
Ask for price |
 POLR3F Antibody, FITC conjugated |
|
1-CSB-PA884440LC01HU |
Cusabio |
|
|
|
|
|
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is FITC conjugated. Tested in the following application: ELISA |
) POLR3F Recombinant Protein (Mouse) |
|
RP163421 |
ABM |
100 ug |
Ask for price |
) POLR3F Recombinant Protein (Human) |
|
RP024124 |
ABM |
100 ug |
Ask for price |
 POLR3F Antibody, Biotin conjugated |
|
1-CSB-PA884440LD01HU |
Cusabio |
|
|
|
|
|
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA |
 (pORF)) Polr3f ORF Vector (Rat) (pORF) |
|
ORF073797 |
ABM |
1.0 ug DNA |
EUR 607.2 |
 (pORF)) POLR3F ORF Vector (Human) (pORF) |
|
ORF008042 |
ABM |
1.0 ug DNA |
EUR 114 |
 (pORF)) Polr3f ORF Vector (Mouse) (pORF) |
|
ORF054475 |
ABM |
1.0 ug DNA |
EUR 607.2 |
 POLR3F Polyclonal Conjugated Antibody |
|
C27655 |
SAB |
100ul |
EUR 476.4 |
 Human POLR3F Protein Lysate 20ug |
|
IHUPOLR3FPLLY20UG |
Innovative research |
each |
EUR 213 |
|
|
|
Description: Human POLR3F Protein Lysate 20ug |
 POLR3F Polyclonal Antibody, HRP Conjugated |
|
A60349 |
EpiGentek |
-
EUR 684.66
-
EUR 302.50
-
EUR 423.50
|
|
 POLR3F Polyclonal Antibody, FITC Conjugated |
|
A60348 |
EpiGentek |
-
EUR 684.66
-
EUR 302.50
-
EUR 423.50
|
|
 POLR3F Polyclonal Antibody, Biotin Conjugated |
|
A60347 |
EpiGentek |
-
EUR 684.66
-
EUR 302.50
-
EUR 423.50
|
|
) Polr3f sgRNA CRISPR Lentivector set (Rat) |
|
K6496101 |
ABM |
3 x 1.0 ug |
EUR 406.8 |
 Polr3f 3'UTR GFP Stable Cell Line |
|
TU266543 |
ABM |
1.0 ml |
Ask for price |
Polr3f 3'UTR GFP Stable Cell Line |
|
TU166697 |
ABM |
1.0 ml |
Ask for price |
 POLR3F 3'UTR GFP Stable Cell Line |
|
TU068441 |
ABM |
1.0 ml |
EUR 1672.8 |
) POLR3F sgRNA CRISPR Lentivector set (Human) |
|
K1685001 |
ABM |
3 x 1.0 ug |
EUR 406.8 |
) Polr3f sgRNA CRISPR Lentivector set (Mouse) |
|
K4059301 |
ABM |
3 x 1.0 ug |
EUR 406.8 |
 (pPM-C-HA)) POLR3F Protein Vector (Rat) (pPM-C-HA) |
|
PV295188 |
ABM |
500 ng |
EUR 723.6 |
 (pPB-C-His)) POLR3F Protein Vector (Rat) (pPB-C-His) |
|
PV295186 |
ABM |
500 ng |
EUR 723.6 |
 (pPB-N-His)) POLR3F Protein Vector (Rat) (pPB-N-His) |
|
PV295187 |
ABM |
500 ng |
EUR 723.6 |
 (pPM-C-His)) POLR3F Protein Vector (Rat) (pPM-C-His) |
|
PV295189 |
ABM |
500 ng |
EUR 723.6 |
 (pPM-C-HA)) POLR3F Protein Vector (Human) (pPM-C-HA) |
|
PV032167 |
ABM |
500 ng |
EUR 394.8 |
 (pPM-C-HA)) POLR3F Protein Vector (Mouse) (pPM-C-HA) |
|
PV217900 |
ABM |
500 ng |
EUR 723.6 |
 (pPB-C-His)) POLR3F Protein Vector (Human) (pPB-C-His) |
|
PV032165 |
ABM |
500 ng |
EUR 394.8 |
 (pPB-N-His)) POLR3F Protein Vector (Human) (pPB-N-His) |
|
PV032166 |
ABM |
500 ng |
EUR 394.8 |
 (pPM-C-His)) POLR3F Protein Vector (Human) (pPM-C-His) |
|
PV032168 |
ABM |
500 ng |
EUR 394.8 |
 (pPB-C-His)) POLR3F Protein Vector (Mouse) (pPB-C-His) |
|
PV217898 |
ABM |
500 ng |
EUR 723.6 |
 (pPB-N-His)) POLR3F Protein Vector (Mouse) (pPB-N-His) |
|
PV217899 |
ABM |
500 ng |
EUR 723.6 |
 (pPM-C-His)) POLR3F Protein Vector (Mouse) (pPM-C-His) |
|
PV217901 |
ABM |
500 ng |
EUR 723.6 |
 Polr3f 3'UTR Luciferase Stable Cell Line |
|
TU216543 |
ABM |
1.0 ml |
Ask for price |
Polr3f 3'UTR Luciferase Stable Cell Line |
|
TU116697 |
ABM |
1.0 ml |
Ask for price |
 POLR3F 3'UTR Luciferase Stable Cell Line |
|
TU018441 |
ABM |
1.0 ml |
EUR 1672.8 |
 Antibody) RNA Polymerase III Subunit F (POLR3F) Antibody |
|
abx236632-100ug |
Abbexa |
100 ug |
EUR 610.8 |
|
|
RNA Polymerase III Subunit F (POLR3F) Antibody |
|
20-abx114612 |
Abbexa |
|
|
|
|
RNA Polymerase III Subunit F (POLR3F) Antibody |
|
20-abx126392 |
Abbexa |
-
EUR 493.20
-
EUR 710.40
-
EUR 218.40
-
EUR 376.80
|
- 100 ul
- 200 ul
- 20 ul
- 50 ul
|
|
|
RNA Polymerase III Subunit F (POLR3F) Antibody |
|
20-abx008320 |
Abbexa |
-
EUR 360.00
-
EUR 526.80
-
EUR 226.80
|
|
|
|
RNA Polymerase III Subunit F (POLR3F) Antibody |
|
20-abx302688 |
Abbexa |
-
EUR 493.20
-
EUR 2214.00
-
EUR 718.80
-
EUR 218.40
-
EUR 360.00
|
- 100 ug
- 1 mg
- 200 ug
- 20 ug
- 50 ug
|
|
|
 (Target 1)) Polr3f sgRNA CRISPR Lentivector (Rat) (Target 1) |
|
K6496102 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 2)) Polr3f sgRNA CRISPR Lentivector (Rat) (Target 2) |
|
K6496103 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 3)) Polr3f sgRNA CRISPR Lentivector (Rat) (Target 3) |
|
K6496104 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 1)) POLR3F sgRNA CRISPR Lentivector (Human) (Target 1) |
|
K1685002 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 2)) POLR3F sgRNA CRISPR Lentivector (Human) (Target 2) |
|
K1685003 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 3)) POLR3F sgRNA CRISPR Lentivector (Human) (Target 3) |
|
K1685004 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 1)) Polr3f sgRNA CRISPR Lentivector (Mouse) (Target 1) |
|
K4059302 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 2)) Polr3f sgRNA CRISPR Lentivector (Mouse) (Target 2) |
|
K4059303 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 (Target 3)) Polr3f sgRNA CRISPR Lentivector (Mouse) (Target 3) |
|
K4059304 |
ABM |
1.0 ug DNA |
EUR 184.8 |
 Recombinant Human POLR3F Protein, His, E.coli-1mg |
|
QP13106-1mg |
EnQuireBio |
1mg |
EUR 3308.4 |
 Recombinant Human POLR3F Protein, His, E.coli-5ug |
|
QP13106-5ug |
EnQuireBio |
5ug |
EUR 186 |
 Recombinant Human POLR3F Protein, His, E.coli-20ug |
|
QP13106-20ug |
EnQuireBio |
20ug |
EUR 241.2 |
 Antibody (HRP)) RNA Polymerase III Subunit F (POLR3F) Antibody (HRP) |
|
20-abx311789 |
Abbexa |
-
EUR 493.20
-
EUR 2214.00
-
EUR 718.80
-
EUR 218.40
-
EUR 360.00
|
- 100 ug
- 1 mg
- 200 ug
- 20 ug
- 50 ug
|
|
|
 Antibody (FITC)) RNA Polymerase III Subunit F (POLR3F) Antibody (FITC) |
|
20-abx311790 |
Abbexa |
-
EUR 493.20
-
EUR 2214.00
-
EUR 718.80
-
EUR 218.40
-
EUR 360.00
|
- 100 ug
- 1 mg
- 200 ug
- 20 ug
- 50 ug
|
|
|
 Antibody (Biotin)) RNA Polymerase III Subunit F (POLR3F) Antibody (Biotin) |
|
20-abx311791 |
Abbexa |
-
EUR 493.20
-
EUR 2214.00
-
EUR 718.80
-
EUR 218.40
-
EUR 360.00
|
- 100 ug
- 1 mg
- 200 ug
- 20 ug
- 50 ug
|
|
|
 ELISA Kit) Human RNA Polymerase III Subunit F (POLR3F) ELISA Kit |
|
abx382365-96tests |
Abbexa |
96 tests |
EUR 1093.2 |
|
|
 pCDH-CMV-MCS-EF1-Puro cDNA Vector cDNA Clon/Exp Vector [pre-packaged virus] |
|
CD510VB-1 |
SBI |
>1 x 10^6 IFUs |
EUR 720 |
 POLR3F Polymerase III Polypeptide F Human Recombinant Protein |
|
PROTQ9H1D9 |
BosterBio |
Regular: 20ug |
EUR 380.4 |
|
Description: POLR3F Human Recombinant produced in E.coli is a single, non-glycosylated polypeptide chain containing 339 amino acids (1-316) and having a molecular mass of 38.1kDa.;POLR3F is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques. |
 pCDF1-MCS1 cDNA Cloning and Expression Vector |
|
CD100A-1 |
SBI |
10 ug |
EUR 483 |
) Polr3f sgRNA CRISPR/Cas9 All-in-One Lentivector set (Rat) |
|
K6496105 |
ABM |
3 x 1.0 ug |
EUR 451.2 |
The info point out that in cells missing RNA-DNA hybrid and Ty1 repressors, elevated ranges of RNA-cDNA hybrids, that are related to duplex nucleic acid constructions, increase Ty1 mobility by way of a Rad52-dependent mechanism. In distinction, in cells missing RNA-DNA hybrid repressors alone, elevated ranges of RNA-cDNA hybrids, that are related to triplex nucleic acid constructions, increase Ty1 mobility by way of a Rad52-independent course of. We suggest that duplex and triplex RNA-DNA hybrids promote transposon mobility by way of Rad52-dependent or -independent mechanisms.
