... or DIO. Lox sites are recognized by the recombinase Cre. Cre first flips the sequence between the first...Lox sites Lox sites, in combination with the Cre recombinase, are used to spatially and temporally control...flipped transgene. By restricting the availability of Cre, lox sites lets researchers control the spatial and... activation of a transgene. Figure 2: FLEX Cre-Lox system.  WPRE The Woodchuck Hepatitis Virus...which aids in nuclear export of mRNA and helps increase the expression of your transgene. The WPRE is ...

...vector from the Gage lab encodes GFP and Cre-recombinase. The Cre-lox system is a technology that can be... Mammalian shRNA Tools for RNAi! Cre-Lox technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral ...vector deposited by Dr. Tyler Jacks. It carries CRE recombinase and a puromycin resistance cassette. No time...the pLKO.1 vector can be found here. Cre-Lox technology Puro.Cre empty vector (Plasmid #17408)   This ...of mouse transgenics. Experimental uses include: Cre-dependent gene expression, gene knockout, and selection...shRNA), mutation/editing (CRISPR-Cas9) or deletion (Cre), together with expression of fluorescent or enzymatic...CRISPR pooled libraries that allow functional screening of thousands of genes in genome-wide experiments...

...instead of using piggyBac™, you could use recombinase technology (Cre/loxP) to remove the donor cassette. While...mutants [7], it is generally not as efficient as Cre recombinase. However, the use of negative selection largely... to DNA toxicity. More importantly, unlike Cre recombinase, the PBx is highly specific, and has not been...applications, including transgenesis, gene-trap screens, and gene editing. How does the piggyBac™ system...of the major uses of the piggyBac™ system is to create stable transgenic cell lines. The traditional method...site-specific nuclease (such as CRISPR/Cas9) to create a double-stranded DNA break. After DNA cleavage... editing? There are primarily two other ways to create specific point mutations by gene editing. First...

...This is achieved through site-specific recombinases such as Cre or Flp that induce DNA recombination at...cells with an inactive opsin gene; in Cre expressing cells, however, Cre induces the recombination of the ...al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents." Scientific reports...recombination. Purification and properties of the Cre recombinase protein." Journal of Biological Chemistry 259.3...loxP and lox511) in opposite orientations. Since Cre does not cause recombination between mismatched recognition...specific neuronal cells? This can be achieved using Cre-dependent viruses, such as the optogenetic FLEx vectors...combination with transgenic animals or rAAVs expressing Cre under a specific cell-type promoter. Once injected...

... PMID: 30678704. Sparse cell labeling using recombinase competition Article contributed by Aliyah Weinstein...Weinstein Sparse Predictive Activity through Recombinase Competition (SPARC) is a fully genetic tool for... Figure 1: Initial steps in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu et al., 2019.  ... Peking University has developed a new CRISPR screening method that makes use of guide RNAs with internal...false-positive rate discovery rate, conventional CRISPR screens require a low multiplicity of infection (MOI) during... annotated human gene, it enables genome-wide screening with a high MOI, which is beneficial for a low... calcineurin dynamics and can serve as a drug-screening platform to identify new antifungals that target...

...type-specific manipulation with GFP-dependent Cre recombinase.” Nat. Neurosci. 18(9) (2015): 1334-1341. PubMed...

... how Cre-lox recombination works. Advances in Cre-lox recombination have included inducible Cre, such...Cai et al. also created an Autobow construct containing both Cre and XFPs. Cre production drives recombination...continued utility of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need a quick...membrane tethered cyan). Without Cre recombination, RFP is expressed. Cre can mediate one of two deletions...construct, OFP (orange), is expressed when Cre is absent; stochastic Cre excision results in one of three recombinations...system employs Cre-mediated deletion, the design of Brainbow-2.1 cleverly combines Cre-mediated deletion...remaining cassette can invert as long as Cre remains expressed; once Cre is removed from the system, the fluorescent...

...It’s an alternative to the conventional Cre or Flp recombinases used to excise markers. It has the advantage...doxycycline, but it can also be regulated spatially with Cre-recombination. Read the paper in Frontiers in Molecular...Cas12a inhibitors Using a bioinformatics and a screening approach, the Jennifer Doudna lab identified three...

...protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be used ...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...OFP (orange) (Livet et al., 2007). Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent...gene directly following the promoter is expressed. Cre-mediated deletion of one or several of these genes...system. One such system is Brainbow 2.0, which uses Cre to trigger the inversion of two genes aligned head-to-head...head-to-head. Another variation, Brainbow 2.1, uses Cre to trigger the inversion of two tandem sets of genes...melanogaster and is under the control of Flp recombinase. The XFPs in Flybow are membrane-tethered, making...

...Versatile Viral Tool for Gene Expression in Mammals Recombinase-based State Machines Enable Order-dependent Logic...fluorophores other than fluorescent proteins can be incredibly useful in FRET experiments. Professor Fessenden's... also really helped put this technology into a concrete context. I recommend this post to anyone wondering...

...fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre, Flp etc.), and tetracycline drug induction...But CRISPRing typical mouse models, including creating Cre-dependent conditional alleles, has remained ...marker, likely by breeding to a mouse line with a recombinase. If it turned out your chimeric mice did not ...plasmid pmScarlet_C1! After creating the T7-DNA template, you'd use ivTRT to create the ssDNA construct as ...CRISPR genome editing has made it easier to create knockout alleles in a variety of species, including...repair template is used. The original design for creating a conditional allele, or floxed mouse, from Yang... and inject the ES cells into a mouse embryo to create chimeric mice. This laborious process takes ~8-...

...genetic tools: activators, inhibitors, sensors, recombinases, and more. With the help of adeno-associated...Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT, with AAV vectors are common approaches... strategies Combining different approaches can increase the specificity of cell targeting even further...delivery of AAV at the injection site results in increased specificity of the targeted cell population.  ...are also developing ways to improve delivery by increasing transgene cargo capacity, enhancing anterograde...

...DreO-bGHpA 121675  AAV9  pAAV-EF1a-fDIO-Cre (FLP-dependent CRE) 55634  AAV1  pAAV-EF1a-mCherry-IRES-... constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-...membrane-embedding voltage detector. Find Archon1 and its cre-dependent version at Addgene! Fluorescent protein...Chimeric channels for neuronal manipulation Recombinases Plasmid Serotype Name 50363  AAV5  AAV ...

...extracted from a sample and subject to isothermal recombinase polymerase amplification (RPA) using a commercially...contains SSB, strand displacement DNA polymerase, recombinase, and the ssDNA-FQ reporters that exhibit fluorescence...based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA per microliter Time...

...CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions, deletions...community members. In addition we’ve experienced incredible growth with our readership expanding from ~10,000...next revolution in biological research. We’re incredibly happy to see that one of our career posts has...protein of interest. 4. Plasmids 101: Blue-White Screening Getting a lot of empty vector contaminants in ...Addgenie Jessica Welch explains how Blue-White screening could be a potential solution. 5. Cpf1: A New ...

...open-reading frame (DIO). Upon coexpression with cre recombinase (either from a mouse line or viral co-injection...specificity dependent on the promoter of the cre recombinase while still taking advantage of the robust ...an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer required...tracing with fluorescent proteins in nontransgenic and cre driver mice. Current protocols in neuroscience / ...: PMC4242517.  7. Madisen L, et al. A toolbox of Cre-dependent optogenetic transgenic mice for light-induced... us to show causality in neural circuits with incredible temporal and spatial precision (I’m not going...optogenetics has already been proven. Therefore, a concrete, well defined, and robust behavioral output is...

...pAAV-hSyn Con/Fon hChR2(H134R)-EYFP that is both Cre and Flp dependent. Plasmid Serotype Name 124603...mCherry)-EGFP 50457 AAV2 pAAV-hSyn-DIO-EGFP Recombinases Plasmid Serotype Name 87306 AAVrg, AAV5...

...made with BioRender.   Grow plasmids in recombinase-deficient strains Since plasmid multimerization...plasmid in a bacterial strain that is deficient in recombinase protein RecA can reduce the frequency of multimerization...generated by SnapGene software.  Thanks to increasing affordability and accessibility, more researchers...by growing seed stocks at 30°C compared to 42°C. Screen multiple colonies to isolate the monomer Sometimes... 1984; Summers et al., 1993). In these cases, screening multiple bacterial colonies using one of the detection...DNA-forming sequences and WRN deficiency independently increase the frequency of base substitution in human cells... self-control and a sense of direction are the secrets of multicopy plasmid stability. Mol Microbiol. ...

...will only be expressed in cells that express cre recombinase. Typically, this feature is used to attain ...Expression of hM4D(Gi)-mCherry from this virus is cre-dependent, meaning that once injected, the DREADD...cell-specific expression in transgenic animals, where cre expression is restricted to a particular cell type...the first time, the Roth Lab recommends verifying cre expression prior to use. The lab additionally recommends...expressing cell lines, which can then be used both for screens (using pooled CRISPR knockout libraries) and individual...Secondly, you can use a LentiCas9-Blast viral prep to create a single stable Cas9-expressing cell line and then...monoclonal Cas9-expression line prior to performing a screen or even a single gene knockout. Top AAV: pAAV-hSyn-DIO-hM4D...

...excised by intentional introduction of the Cre recombinase. This is a time-tested system but does require...control when you turn off expression for good, the Cre-Lox system offers inducible yet permanent gene excision...some permanent knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for ...!  Turn it on Turn it on forever Permanently increasing or turning on expression of a new protein is ... gRNA, to your gene of interest to temporarily increase expression. There are several flavors of activators...