The lentiviral transduction particles are produced from an shRNA lentiviral non-target control plasmid. (sigmaaldrich.com)
PPLK/GFP+PURO-ATG4B SHRNA-1 PLASMID - 1 kit is backordered and will ship as soon as it is back in stock. (dnamethsoc.com)
A novel non-viral vector, G5-BGG, was synthesized and formed into a gene complex with shRNA plasmid. (physiciansweekly.com)
shRNAs are delivered as 3 individual constructs of 5 µg purified plasmid and a separate scrambled control plasmid. (genecopoeia.com)
Functional assays by a reporter gene/protein - Alternatively, when the endogenous transcript level of gene of interest is low and/or when qRT-PCR and western blot analysis is not feasible or possible, KD effect can be measured by co-transduction of shRNA clone with an expression clone plasmid which is transcribed into a chimeric mRNA transcript consisting of a reporter gene and the target gene ORF. (genecopoeia.com)
At Altogen Labs we specialize in the shRNA design and synthesis of shRNA-encoding vector constructs as well as the purification of shRNA encoding plasmid DNA and transfection of shRNA-expressing constructs into cultured cells. (altogenlabs.com)
You can easily produce a recombinant lentivirus particle in HEK293T or HEK293FT cell line in high titer using GeneMedi's lentivirus Vector System, including the lentivirus expression plasmids (overexpression, shRNA or Crispr-cas9-gRNA), the envelope protein VSV-G expressing plasmid pMD2G and the packaging plasmid pSPAX2. (genemedi.net)
Vectors8
You can select shRNA vectors with different antibiotic selection markers or fluorescent-antibiotic fusion dual markers. (gentarget.com)
When you design shRNA vectors on VectorBuilder's online platform, you will have the option to search for your target genes in our database. (vectorbuilder.kr)
Our Lentivirus IPTG-Inducible shRNA Knockdown vectors are derived from the third-generation lentiviral vector system. (vectorbuilder.kr)
A) Lentiviral vectors carrying IPTG-inducible U6-based scramble or EGFP-targeting shRNA expression cassettes were packaged into the corresponding lentiviral particles and transduced into HEK293T cells stably expressing EGFP. (vectorbuilder.kr)
Lentiviral vectors can be used to deliver shRNAs, thereby providing the ability to infect most mammalian cell types with high efficiency, regardless of proliferation state. (arizona.edu)
Lentiviral inducible shRNA vectors are readily available, but unfortunately the process of cloning, screening, and testing shRNAs can be time-consuming and expensive. (arizona.edu)
Furthermore, we provide a detailed guide for utilizing these vectors, including shRNA design strategy and simplified screening methods. (arizona.edu)
Lastly, we display the robustness of the system with a doxycycline titration and recovery time course and provide a cost/benefit analysis comparing our system with purchasing pre-designed shRNA vectors. (arizona.edu)
Knockdown18
Embryonic knockdown of the function of homologs of these genes in rat neocortical projection cell progenitors by in utero electroporation of plasmids encoding small hairpin RNA (shRNA) revealed that all three genes disrupted neuronal migration to the neocortex. (nih.gov)
This shRNA non-mammalian control was designed using our Turbo GFP sequence and may cause some knockdown of tGFP. (sigmaaldrich.com)
When conducting experiments using MISSION ® shRNA clones, the proper controls should be a key element of your experimental design to allow for accurate interpretation of knockdown results. (sigmaaldrich.com)
shRNA of varying lengths (19 to 29 bases) was designed using a proprietary algorithm to make shRNA expression constructs that have high knockdown efficiency with minimal off-target effect. (genecopoeia.com)
shRNA is driven by pol III promoters (H1, U6) or pol II promoters (CMV, EF1a, CAG and CBh), both with great knockdown efficiency. (genecopoeia.com)
Functional assays - The shRNA knockdown effect can be studied by end-point biological and biochemical assays such as cell proliferation, colony formation, cell cycle analysis, and migration based on the potential role played by each gene /protein. (genecopoeia.com)
RNAi knockdown can be introduced by synthetic short double-strand RNA ( siRNA ) or vector-expressed stem-hairpin RNA ( shRNA ) which is further processed by the Dicer enzyme to produce double-stranded short RNAs. (gentarget.com)
The shRNA lentivector can be used directly in transfection for transient knockdown or to produce lentivirus for long-term knockdown experiments. (gentarget.com)
This shRNA lentivirus serve as a useful negative control / reference for interpretation of target specific knockdown results. (gentarget.com)
The Lentivirus IPTG-Inducible shRNA Knockdown vector is a highly efficient system for achieving temporal knockdown of target genes in a wide variety of mammalian cell lines and offers an effective tool for studying genes essential for cell development or cell survival. (vectorbuilder.kr)
Our Lentivirus IPTG-Inducible shRNA Knockdown vector contains a LacI sequence and a modified human U6 promoter with two repeats of the LacO element (U6/2xLacO). (vectorbuilder.kr)
Our database ranks all available shRNAs for a target gene in order of their decreasing knockdown scores and recommends testing the top 3 shRNAs with the highest knockdown scores. (vectorbuilder.kr)
Our shRNA stem-loop sequences are optimized to mediate efficient shRNA processing and target gene knockdown. (vectorbuilder.kr)
Our lentivirus IPTG-inducible shRNA knockdown vector has been validated for highly efficient target gene knockdown in the presence of IPTG as shown in Figure 1 below. (vectorbuilder.kr)
Through mechanisms of mRNA cleavage when in complex with the RISC complex, shRNA can be used to knockdown the expression of target proteins. (altogenlabs.com)
Silencing of TCOF1 expression (shRNA) radiosensitized progenitor cells, a response conserved in human cells with TCOF1 knockdown. (cdc.gov)
Promoter12
Our HIV-based pGreenPuro (EF1α) shRNA Cloning and Expression Lentivector shRNA Cloning and Expression Lentivector drives expression of your shRNA template from the H1 promoter, and after processing in the cell, your shRNA will be converted into siRNA. (systembio.com)
After packaging and transduction, the vector will integrate into the genome and your shRNA will be transcribed from the H1 promoter using RNA polymerase III. (systembio.com)
Expression cassettes of all shRNA clones are fully sequenced including the promoter, sense and antisense target sequences, hairpin, terminator, and other linker sequences. (genecopoeia.com)
GenTarget provides easy-to-use, unique, advanced cloning kits for constructing shRNA lentivectors using either the human U6 promoter or the human H1 promoter . (gentarget.com)
Unless you already know which promoter is better in your specific cell type, you can select either promoter for shRNA expression. (gentarget.com)
Each shRNA construct is driven by the U6 promoter and contains a GFP reporter. (criver.com)
The shRNA targeting the gene of interest (GOI) is placed downstream of the modified U6 promoter with an additional LacO sequence located downstream of the shRNA. (vectorbuilder.kr)
In the presence of IPTG, LacI undergoes a conformational change and is no longer able to bind to LacO, thereby allowing the shRNA to be transcribed by the U6 promoter. (vectorbuilder.kr)
The modified human U6 promoter with two LacO elements drives high-level transcription of the downstream shRNA in mammalian cells in the presence of IPTG, which prevents LacI from binding to LacO. (vectorbuilder.kr)
The shRNA expression is driven by an U6 promoter. (vectorbiolabs.com)
Deep sequencing of transfected cells showed accurate processing of individual shRNA-miRs and that their expression did not decrease with the distance from the promoter. (utmb.edu)
This AAV expresses a scramble shRNA sequence under the control of U6 promoter, with the GFP co-expression under a CMV promoter. (vectorbiolabs.com)
Lentivirus5
shRNAs that generate siRNAs intracellularly are expressed from amphotropic lentivirus viral particles, allowing screening in a wide range of mammalian cell lines. (sigmaaldrich.com)
The human pancreatic cancer Capan-1 cell line was first infected with lentivirus-mediated shRNA and then analyzed by real-time PCR, and the results were further confirmed by Western blotting. (academicjournals.org)
DcR3 protein in our experimental results showed significant expression among the pancreatic cancer cell lines and their decreased expression by lentivirus-mediated shRNA resulted in the inhibition of cell proliferation and augmentation of apoptosis. (academicjournals.org)
Lentivirus and AAV for shRNA are also available. (genecopoeia.com)
The shRNA lentivirus can be transduced into dividing and non-dividing cells and into the majority of mammalian cell types. (gentarget.com)
Genes8
OmicsLink ™ shRNA clone collections include lentiviral and non-viral vector-based shRNA constructs against genome-wide human, mouse and rat genes. (genecopoeia.com)
By arraying the shRNA clones of known pathway(s) in 96 or 384 well plates, the role for a group of genes can be studied in a pathway. (genecopoeia.com)
Validation studies - The OmicsLink™ expression ready ORF cDNA clones together with the same ORF clones that contain silent mutations in shRNA target sequence regions can be used for shRNA validation studies and gene/protein functional rescue studies for genes/proteins targeted by corresponding shRNA. (genecopoeia.com)
Negative shRNA controls containing a insert that designed has no homogous to any human or mouse transcripts (should not target any known human or mouse genes). (gentarget.com)
Short hairpin RNA (shRNA) refers to a sequence of RNA that has the ability to silence genes in a sequence-specific manner via RNA interference (RNAi). (altogenlabs.com)
Module 3 of Cellecta´s Human Genome-Wide (hGW) shRNA Library targeting ca. 6,300 genes, including cell surface markers, extracellular matrix genes, and DNA binding genes. (nucleusbiotech.com)
Cellecta´s 3-module pooled lentiviral Human Genome-wide shRNA Library cloned in pRSI16cb targets nearly all protein-encoding genes of the human genome, and each gene is targeted by 8 hairpins. (nucleusbiotech.com)
Multiplexed miRNA-based shRNAs (shRNA-miRs) could have wide potential to simultaneously suppress multiple genes. (utmb.edu)
Short hai1
Background: Short hairpin RNA (shRNA) is an established and effective tool for stable knock down of gene expression. (arizona.edu)
RNAi3
Small interfering RNAs (siRNAs) expressed from short hairpin RNAs (shRNAs) are a powerful way to mediate gene specific RNA interference (RNAi) in mammalian cells. (sigmaaldrich.com)
To produce siRNAs for RNAi using the pSIF1-H1-H2Kk Cloning and Expression Lentivector, first clone your shRNA template into the unique BamHI or EcoRI sites in the vector. (systembio.com)
For shRNA design we apply rules like those used by the RNAi consortium. (vectorbuilder.kr)
SiRNA3
The shRNA is transcribed as a single strand with a sense-loop-anti-sense structure that folds into a hairpin, and is then processed by DICER to produce an active siRNA molecule (Figure 1). (systembio.com)
Generating siRNA from the pGreenPuro (EF1α) shRNA Cloning and Expression Lentivector. (systembio.com)
VIRsiRNAdb is a database of siRNA/shRNA targeting viral genome regions. (wikipedia.org)
Clones2
In these cell lines, MISSION shRNA clones permit rapid, cost efficient loss-of-function and genetic interaction screens. (sigmaaldrich.com)
Single gene down-regulation - The knocking down (KD) effect of the shRNA clones for a single gene can be studied and compared with that of a scrambled nucleotide control clone which is included for free with every shRNA clone order. (genecopoeia.com)
Hairpin2
Previously, AAV9-mediated delivery of a small hairpin RNA (shRNA) targeting human SOD1 resulted in efficient downregulation in mice and non-human primates (NHPs). (neurology.org)
Altogen Labs offers a complete service for preparing DNA templates and synthesizing hairpin shRNAs. (altogenlabs.com)
Particles1
Unlike murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the specific shRNA construct into differentiated and non-dividing cells, such as neurons and dendritic cells, 1 overcoming low transfection and integration difficulties when using these cell lines. (sigmaaldrich.com)
Constructs2
By default, shRNA clone constructs are designed to target the splice variant (accession number) you selected for this gene WITHOUT consideration of other possible variants of this gene. (genecopoeia.com)
Four pre-designed shRNA constructs targeting golgi-associated, gamma adaptin ear containing, ARF binding protein 1 (GGA1), transcript variant 5 and one scrambled control. (criver.com)
VectorBuilder1
VectorBuilder has created shRNA databases that contain optimized shRNAs for common species. (vectorbuilder.kr)
Sequence6
And you can find all shRNA lentivector sequence here. (gentarget.com)
Upon entering your gene name, you will see detailed information on all shRNAs against your GOI available in our database, including a link to UCSC Genome Browser to view these shRNAs in the context of genomic sequence and all the transcript isoforms. (vectorbuilder.kr)
The library includes shRNA-specific barcodes that provide an alternative to direct sequencing of the shRNA target sequence which is difficult to sequence due to the stem-loop structure. (nucleusbiotech.com)
We found that a sequence of 30 nucleotides flanking the miRNA duplex was sufficient for efficient processing of shRNA-miRs. (utmb.edu)
We inserted multiple shRNAs in tandem, each containing minimal flanking sequence from a different miRNA. (utmb.edu)
Results: Notably, we emphasize the importance of loop sequence design and demonstrate that the addition of a single mismatch in the loop stem can greatly improve shRNA efficiency. (arizona.edu)
Transfection2
We found that age of transfection did not affect the percentage of unmigrated neurons--transfection with Kiaa0319 shRNA resulted in heterotopia formation at all three ages. (nih.gov)
Lentiviral and non-viral vector options allow the transfection or transduction of shRNA into difficult-to-transfect cells as well as more conventional cell lines. (genecopoeia.com)
Inducible2
Furthermore, the use of inducible promoters to drive shRNA expression allows for more thorough investigations into the specific timing of gene function in a variety of cellular processes. (arizona.edu)
Lentiviral vector-mediated doxycycline-inducible USP39 shRNA or cDNA expression in triple-negative breast cancer cells. (arizona.edu)
Lentivector1
We also can do the shRNA cloning and virus works for you, providing shRNA lentivector cloning services with the fastest turnaround time and the best prices. (gentarget.com)
Viral1
This was efficiently packaged into an AAV9 viral capsid, creating a new AAV9-SOD1-shRNA ("AVXS-301" herein) translatable to the clinic. (neurology.org)
Human3
To develop a first-in-human gene therapy for amyotrophic lateral sclerosis (ALS) using intrathecal AAV9-SOD1-shRNA administration. (neurology.org)
We modified the experimental vector for clinical trials using an expression cassette for the delivery of shRNA against human SOD1 devoid of any foreign transgenes (GFP) but maintaining the same cassette size. (neurology.org)
Here, we employed a human pancreatic differentiation platform complemented with an shRNA screen in human pluripotent stem cells (PSCs) to identify potential drivers of early endoderm and pancreatic development. (mdpi.com)
Cloning2
shRNA Cloning and Expression Lentivectors. (systembio.com)
Conclusions: Our aim was twofold: first, to take a very useful shRNA vector and make it more amenable for molecular cloning and, secondly, to provide a streamlined protocol and rationale for cost-effective design, cloning, and screening of shRNAs. (arizona.edu)
MRNA2
qRT-PCR - Targeting of the corresponding mRNA by a shRNA expression cassette leads to cleavage and degradation of the mRNA and possible subsequent reduction of protein expression. (genecopoeia.com)
The destruction of the target gene mRNA by targeting shRNA results in the degradation of the reporter gene and, therefore, translation reduction of reporter protein enzyme. (genecopoeia.com)
Targeting3
Addgene: shRNA targeting alpha-synuclein prevents neurodegeneration in a Parkinson's disease model. (addgene.org)
We used this system to express one shRNA-miR targeting CCR5 and six shRNA-miRs targeting the HIV-1 genome. (utmb.edu)
shRNAs Targeting a Common KCNQ1 Variant Could Alleviate Long-QT1 Disease Severity by Inhibiting a Mutant Allele. (cdc.gov)
Neurons1
Overmigration of neurons transfected with Dcdc2 shRNA, while present following transfections at the later ages, did not occur following E14.5 transfections. (nih.gov)
Puromycin1
Puromycin-resistant lentiviral control shRNA vector, pLKO.1 induces unexpected cellular differentiation of P19 embryonic stem cells. (arizona.edu)
Protein1
We transfected litters with Dcdc2 shRNA, Dyx1c1 shRNA, Kiaa0319 shRNA, or fluorescent protein (as a control) at each of three gestational ages (E14.5, E15.5, or E16.5). (nih.gov)
Sequences1
Here, we describe a simple strategy to express a large number of shRNA-miRs using minimal flanking sequences from multiple endogenous miRNAs. (utmb.edu)
Target1
It is useful as a negative control in experiments with the MISSION shRNA target sets. (sigmaaldrich.com)
Algorithm2
shRNAs are designed using the SplashRNA algorithm. (mskcc.org)
We embarked on collaborative work with fellow MSK researchers Scott Lowe and Cristina Leslie to develop a new sequential classification algorithm a sequential classifier (SplashRNA)1, to predict potent microRNA-based shRNAs. (mskcc.org)
Unlike1
Unlike one shRNA-miR, the seven shRNA-miR transduced T cells nearly abrogated HIV-1 infection in vitro. (utmb.edu)
Proteins1
This system utilizes the interaction between LacI (repressor) and LacO (operator) proteins, derived from the bacterial lactose operon to regulate shRNA expression in the presence and absence of the lactose analogue, isopropyl-ß-D-thiogalactoside (IPTG). (vectorbuilder.kr)