...research and many different constructs are available to the community. Each construct has unique features, such...RNA (hpRNA). This method is facilitated by DNA constructs that enable insertion of ~400 bp complementary...sequence (discussed below). Examples of these constructs include pGAPi, which allows easy creation of ... address these shortcomings, both short hpRNAs (shRNAs) and artificial micro RNAs (amiRNAs) are frequently...precursors of microRNAs found in the genome, whereas shRNAs are exogenously derived. Importantly, instead of...single ~21-nt interfering RNA is produced using shRNA/amiRNA. Although sh- and amiRNAs are likely more...Arabidopsis species, respectively. Short hairpin RNA (shRNA) is conceptually similar to the microRNA precursor...

...inducible gene overexpression, knockdown (shRNA, miR-30-shRNA), mutation/editing (CRISPR-Cas9) or deletion...vectors can regulate gene expression by delivering shRNA or miRNA. The ever expanding CRISPR field makes ...in mammalian cells.  Learn more about Mammalian shRNA Tools for RNAi! Cre-Lox technology CAG-GFP-IRES-...empty backbones) here. Cloning and expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has...Genetic Perturbation Platform, to produce their shRNA library to support functional investigations of ...sequence that can be removed and replaced with an shRNA sequence of your choice. More information and a ...This modular and flexible system can be used to construct complex lentiviruses for modification of mammalian...

...several options to allow you to do exactly that: shRNAs are short RNA sequences which target complementary...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...selectable marker. This vector can express your construct in target cells transiently or constitutively ...the viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA or a sgRNA...Some customization is required, but many viral constructs have already been built and are available on ... embryonic stem cells. Viral integration of a construct is also reliably higher than traditional transfection...

...item of 2005. shRNA - In 2006, we saw an increase in popularity of empty backbones for shRNA expression,...plasmids you'll note that those are lentiviral constructs. In 2007 we continued to see requests grow for...beta-catenin reporter M50 Super 8x TOPFlash, scramble shRNA for use as a negative control, and empty backbones...

...sgRNA (U6) expression, shRNA/shRNA-miR30 constitutive (H1, U6, 7SK) or inducible shRNA-miR30 (CMV-TO) expression...expression, or a promoterless construct. Pre-constructed Entry vectors containing Cas9, EGFP, mCherry, iRFP...constitutive or inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene...of inducible promoter constructs. Up to 34 building blocks were used to construct 20kb long inserts. The...att sites are supplied as individual reporter constructs or as the fully assembled pMemoryArray.   Yang...Biotechnol Lett. 2015. 37:383-389. Toolkit for constructing fusion proteins with customizable N- and C-terminal... to 5 different elements. Lentiviral vectors constructed with the MuLE toolbox have been used for a variety...

...an RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters. This...commonly employed in lentiviral and retroviral constructs and we will discuss these in a future post on...expression in plants. H1 small RNA expression shRNA From the human polymerase III RNA promoter Constitutive...expression in neuronal cells. U6 small RNA expression shRNA From the human U6 small nuclear promoter Constitutive...

... rely on the random integration of a reporter construct in close proximity to an endogenous gene. Each...detail below), but fundamentally the reporter construct monitors the expression of a nearby gene. The ...an uncharacterized gene. The actual reporter construct can be something phenotypic (such as a fluorescent...the expression of the gene (by CRISPR knockout, shRNA depletion, etc.) and ask what the phenotype is without...monitor. Gene traps Gene traps are promotor-less constructs that contain splice acceptors. They require insertion...type of trap, there are many ways to insert your construct to generate a collection of monitored genes and...

...targeting system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder...endogenous RNA imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head for gene expression... 36690762.   Heat-inducible lentiviral CAR constructs by: Brian O'Neill  A clever system for reducing...system is a set of heat-inducible lentiviral CAR constructs, the most basic of which is depicted here: ...days - after just 15 minutes of FUS.  Similar constructs to the one above can be made for expressing your...

...assembled into a final transgenesis construct. Plasmids constructed using Golden GATEway cloning have been...complexity of assembling recombination of transgenesis constructs? A new cloning system combining the advantages...simplifies the cloning process for complex DNA constructs, particular for those involving recombination...Multisite Gateway cloning to generate the final construct. Up to eight entry vectors can be used for ...methods, particularly for complex transgenesis constructs. Kirchmaier et al., PLOS One 2013 Oct 7;8(10...The Janovjak lab has deposited their 3 Opto-RTK constructs – Opto-mFGFR1, Opto-hEGFR, and Opto-hRET – as...as well as various LOV-domain-mVenus constructs used in the study for those who would like to extend this...

...typically an expression construct, to cells/organisms. This is often cDNA, shRNA, or something similar,...), it’s still used to deliver gene expression constructs. Sandwiched between two inverted terminal repeats...just like AAV, which encompass the expression construct and promoter that you wish to deliver to your ...plasmids in our collection. Want to make a vector construct on your own? No problem, we are here to help with...

...shrink the construct. They identified ADAR2DD(delta984-1090), which decreases the REPAIRv1 construct size from... A->G editing, they employed hyperactive ADAR constructs, like ADAR2DD(E488Q). They also adjusted the ...Although REPAIRv1 knockdown is more precise than shRNA knockdown, it still displays substantial off-target...binding to decrease off-targets. Of the tested constructs, ADAR2DD(E488Q/T375G) (REPAIRv2) had the highest...dRanCas13b truncations that would allow packaging the construct for viral delivery. They found C-terminal truncations...

...suppresses neuronal transmission within minutes. Constructs using the photodimerizer system CRY2-CIB1 to ...response has been a longstanding challenge. Using shRNA, TALEN, or ZFN to edit immune cells poses challenges...immune genes into pXPR_053. They expressed this construct in bone marrow stem cells from Cas9-expressing...

...of viral constructs is a useful tool as it provides a means to quickly check that a construct is functional...variety of applications including cDNA expression, shRNA-mediated knockdown, Tet and Cre-regulated expression...making a stable cell line. When expressing a viral construct transiently care needs to be taken to ensure that...

...transfer plasmid encoding a transgene, sgRNA, or shRNA of interest: The transgene sequence is flanked by...wildtype LTR promoter is no longer present in these constructs. Why do many γ-retroviral plasmids have similar...

...integrated by the virus. If an oncogene overexpression construct is being delivered, that is a higher risk cargo... GFP. Similarly, if the viral vector carries an shRNA to a gene essential for cellular function, this ...

...AAV vectors to deliver chemogenetic/optogenetic constructs into our cells of interest. Although they have...time until expression, it is possible to use AAV constructs with self-complementing structure (scAAV) (7)... would result in cell death. With several new constructs becoming available, AAVs could be much better...such as microRNAs (miRNAs), short hairpin RNAs (shRNAs), antisense oligonucleotides (ASOs), zinc finger...

...the different vectors used, lentiviral vector constructs have proven very useful due to their ability ...lentiviral vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors...

...Howarth lab created the MBPx-SpyCatcher construct. This construct is used to produce a maltose binding protein...However, the laborious construction of repair templates and guide RNA constructs, as well as the difficult... Human Kinase Domain Constructs Kit, a library of human kinase domain constructs developed through a large-scale... that results in the creation of many E. coli constructs with greatly varied levels of expression. The...can be used to easily create protein expression constructs fused with epitopes or fluorophores, generate...platform for generating both simple and complex DNA constructs Have you ever wished you could design a gene...overexpression experiment, generate the necessary constructs, and complete screening in just 3 days? Thanks...