...efficiency of Cre recombination varies for different constructs and cell types. Second, Cre may induce recombination...expression of Cre from the standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor... using multiple recombinases. To make multiple, precise genome edits, the recombinases used must have ...stop sequence. In contrast, Dre recombinase, the close relative of Cre, displayed an excision efficiency...determine the cross-reactivity of these recombinases, each recombinase and reporter construct pair were tested...however, the other recombinase/reporter pairs were inactive, even at very high recombinase levels. These data...toxicity of these recombinases by crossing flies homozygous for a UAS-driven recombinase with flies heterozygous...

...under the control of Flp recombinase together with inducible Cre recombinase (CreERT2), visualizes individual...the control of Cre recombinase are used in combination with mouse lines that express Cre only in the cell...Katrin Michel. Cre-lox is an incredibly popular and powerful site specific recombinase (SSR) system, but...LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2 #14797). These Cre/ FLP versions are retained in the...control without modification - either Cre is there or it’s not. Cre-mediated possibilities for site specific...control FLP and Cre expression have been developed. Read on to learn more about FLP-FRT, Cre-lox, and how... works in a similar fashion to the Cre system. The FLP recombinase recognizes FRT sites and initiates ...

...AAV-GfaABC1D-Cre-4x6T AAV5 Recombinases Stanley Thomas Carmichael New viral prep pAAV-EF1a-fDIO-Cre AAV5 ...AAV5 Recombinases Esteban Engel New serotype pAAV-hSyn-Cre-P2A-dTomato AAV PHP.eB Recombinases Rylan Larsen...Collection. These preps drive expression of either Cre recombinase or a fluorescent marker in the various cell...- pAAV-AiE0450h-minBG-iCre(R297T)-BGHpA (Alias: CN3778) AAV PHP.eB Recombinases Jonathan Ting New viral...pAAV-AiE0779m_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN3779) AAV PHP.eB Recombinases Jonathan Ting New viral...- pAAV-AiE0452h-minBG-iCre(R297T)-BGHpA (Alias: CN3781) AAV PHP.eB Recombinases Jonathan Ting New viral...

...expression of other recombinases. Multiple orthogonal attB-attP pairs per recombinase may be arranged on...different recombinases are overlapping or nested on a register, then the operation of one recombinase on the...they can control recombinase activity. For example, post-transcriptional recombinase activation via RNA...reduces the content of our paper, “Synthetic recombinase-based state machines in living cells” (1), into...explanation of how to adapt our engineered devices, recombinase-based state machines for your own experimental... about the order or timing of these factors. Recombinase-based state machines (RSMs), which take on a ...drives expression of a distinct large serine recombinase that recognizes its own attB and attP recognition...

...have created a cassette that will express GeneR only in the presence of Cre. In this example, Cre would...orientation of the flanking loxP sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment...orientation specificity of the site-specific recombinases (SSRs) Cre and FLP. SSRs bind DNA at target sites ...induce site specific recombination events: Cre recombinase binds loxP sites, while FLP binds FRT sites...and a pair of FRT sites and include both Cre and FLP recombinases.   Figure 1 (modified from Schnutgen... absence of, or mCherry in the presence of Cre recombinase.   Once the SSR of choice is introduced...events are impossible even in the presence of Cre recombinase. This plasmid now specifically drives expression...

...tissue-specific cell types: Cre-lox recombination. What is Cre-lox? The Cre-lox system is a technology ...marker, Cre can be used to easily perform this eviction. Regulated Cre expression – placing Cre downstream...by making the Cre inducible (such as with tamoxifen or doxycycline), the Cre recombinase can be expressed...constructed. Additionally, Cre-containing adenovirus (Ad-Cre) or AAV (AAV-pgk-Cre) has been used to successfully...components derived from the P1 bacteriophage: the Cre recombinase and a loxP recognition site. The P1 bacteriophage...components for use in genome manipulation. Cre recombinase, originally named because it “causes recombination...How does it work? As alluded to above, the Cre recombinase catalyzes the site specific recombination event...

...models that express recombinases like Cre or Flp in their cells of interest and recombinase-dependent AAV vectors...design Recombinase-dependent vectors utilize pairs of specific sequences, lox (recognized by Cre) or frt...this spurious expression?   Figure 1.  Cre-recombinase binds to either pair of lox sites, let's say... On the left homologous pairs of scrambled cre-recombinase sites lead to greater frequency of reversion...system is off-target or spurious expression of recombinase-dependent AAV vectors. In this article, we will...excise a gene that is located between them. Early recombinase-dependent vector designs used a lox-STOP-lox ... to prevent transcription in cells where the recombinase was not present (Kuhlman, SK. 2008). While theoretically...

... as a Cre-dependent Flp-expressing AAV. See our recombinase AAV inventory here New recombinases Plasmid... EGFP Recombinase AAV Finally, we are excited to add Flp-expression AAV to our recombinase AAV inventory...inventory. Flp is another useful recombinase that can be used in combination with Cre for intersectional control... µL size serotype testing) pAAV-hSyn-EGFP Recombinases Plasmid Serotype Name 55634  AAV1  pAAV-EF1a-mCherry-IRES-Flpo...

...inventory of recombinases and recombinase-dependent vectors. We recently released several recombinases in various...Flp-dependent Cre AAV in AAV9 and AAVrg. See our new additions here: Cre vectors pAAV-EF1a-fDIO-Cre (121675...expression. This is a Cre-dependent switch that expresses nuclear mCherry in the absence of Cre, and expresses...Calcium sensors and GABA sensors Optogenetics Recombinases And more! Control AAV For many AAV experiments...expresses nuclear EGFP in the presence of Cre. Biosensor AAV (calcium sensors and GABA sensors) Calcium sensors...sensor (see below), in addition to our inventory of Cre-dependent GABA sensors. See all our GABA sensor AAV...pAAV-CamKIIa-ChrimsonR-mScarlet-KV2.1 (124651-AAV9) Recombinase AAV Targeting and manipulating specific, genetically-defined...

...C-terminal and N-terminal Cre fragments together and permitting recombinase activity. Opto-Cre-Vvd is precise, ... inducible Cre for precise control of bacterial gene expression By Meghan Rego Recombinases are enzymes...RFP reporter plasmid for recombinase activity were exposed to blue light. Cre-Vvd showed substantially...Mary Dunlop’s lab generated a photosensitive Cre, Opto-Cre-Vvd, by splitting and joining the protein regions...light. Unlike other split-Cre systems which can take up to 24 hours to cut, Opto-Cre-Vvd is fast and can cut...pAAV-hSyn-Cre-P2A-dTomato is now available in AAV1 and AAV8, as well as AAV5 and AAVrg. Find the Cre-expressing...improved activity as compared to other recombinase/photodimer pairs. Created with BioRender.com In their ...

...(Gq)-mCherry Bryan Roth   Recombinases Recombinase systems such as Cre-lox allow for more accurate ...types, recombinases allow you to further restrict gene expression to areas in which the recombinase and ...coexpressed. Three new recombinases from the Cepko Lab are now available as AAV1. CRE-DOG constructs 69570...dependent Cre. The two segments come together in the presence of GFP to form a functional Cre enzyme that...ChR2 for targeted expression DREADDs GFP dependent Cre and FLPo And more!  New lentiviral preps SARS-...control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain...future therapies. Our popular plasmid encoding a Cre dependent version of the DREADD hM3D is now also ...

....GFP-Cre.WPRE.SV40 AAVrg Recombinase James M. Wilson New serotype pENN.AAV.CamKII 0.4.Cre.SV40 AAVrg...pAAV-syn-Flpo AAVrg Recombinase Ian Wickersham New viral prep pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 AAV2 Recombinase...James M. Wilson New serotype pENN.AAV.hSyn.Cre.WPRE.hGH AAV2 Recombinase James M. Wilson New serotype ...pAAV-AiE0441h_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN4825) AAV-PHPeB Recombinase Jonathan Ting New viral...AAVrg Recombinase James M. Wilson New serotype AiP12237 - pAAV-AiE0452h-minBG-SYFP2-WPRE3-BGHpA (Alias:...

...However, the Cre-dependent AAV cargo would only become activated in the presence of Cre recombinase, which ...by injecting a Cre-dependent AAV into Broca’s area of a mouse specifically expressing Cre in the hippocampus...phenomenon is to use a retrograde recombinase-expressing vector (e.g. Cre or FLP) to transduce the projection...genetically controlled function. By using AAVrg in Cre transgenic mouse lines, you can access and manipulate...hippocampus (Fig. 3). The Cre-dependent AAV would access the neurons (Fig. 3A) that project to Broca’s area...titers for efficient retrograde transport. For recombinase-dependent vectors, this can cause challenges ...projection pathway neurons and then inject a recombinase-dependent chemogenetic vector directly into the region...

... in recombinase-dependent sequences. Recombinase-dependent sequences may be subject to recombinase-independent...pre-recombined population that does not require the recombinase for transgene expression. Once packaged in AAV...pre-recombined DNA could lead to transgene expression in recombinase-negative animals, producing misleading results... hundreds of viral vectors and estimate that recombinase-independent recombination occurs in 0.1-0.6% ...transgene expression but limits the likelihood of recombinase-independent expression. Next-generation sequencing...parallel because the risk of accidental errors increases. In addition to human error, fragments of DNA ...to the reference sequence of the plasmid used to create the viral prep. In a clean viral prep, like AAV...

...transcription or Cre recombinase activity only in the presence of GFP. These systems, known as T-DDOG and Cre-DOG,...Although Cre isn’t a modular protein, Tang et. al were able to create a split version of Cre that is only...Podcast Interview with Connie Cepko Combining Cre and GFP with Cre-DOGs The T-DDOG results showed that GFP ...started with another biological workhorse, Cre recombinase, which induces recombination at LoxP sites.... of interest blocks transcription when Cre is absent. When Cre is present, it removes the STOP cassette...when dimerized by the GBPs/GFP. The new system, CRE-DOG (Cre Dependent On GFP), is activated by GFP and derivatives... cord. Cre-DOG is also suitable for optogenetic studies, and since cells infected with AAV-Cre-DOG retain...

...site-specific serine recombinase. First, the prime editor installs the recombinase “landing sites,” then...then the recombinase handles the installation of the donor sequence. The latest work evolves and engineers...engineers Bxb1 recombinase variants to optimize PASSIGE efficiency (Pandey et al., 2024). The best-performing...editing installs two sequences containing attB/P recombinase landing sites on opposite DNA strands. When the...edited sequence with the attB/P landing site. A recombinase integrates a sequence from a donor plasmid containing... in mammalian cells via continuously evolved recombinases and prime editing. Nature Biomedical Engineering...A., Tao, Y. A., Duby, J. E., Steinbeck, B. J., McCreary, J., Pierce, S. E., … & Liu, D. R. (2024). Efficient...

...limiting amount of AAV are injected into the brain of Cre-expressing mice to trigger recombination and expression... 2019. Sparse Predictive Activity through Recombinase Competition (SPARC), a genetic tool that allows...expressed and the cells are not labeled. PhiC31, a recombinase. It specifically integrates an attP site into...variety of fly cell types. Because the PhiC31 recombinase can work in other model organisms including mouse...site upstream of the stop cassette results in decreased efficiency of recombination and preferentially...

...encoding Cre recombinase under the control of the synapsin promoter. It can be used to drive Cre expression...be to inject your Cre-dependent mCherry mouse with the pmSyn-EBFP-Cre AAV. Because Cre expression is controlled... could breed your Cre-dependent mCherry line with a separate line that expresses Cre in neurons, but both...expression of any Cre-dependent gene of interest in neurons. This ability to be paired with any Cre-controlled...interest makes pmSyn-BFP-Cre highly modular. Further uses of pmSyn1-EBFP-Cre Of course there are also ... cre-recombinase...AAV that became available in 2017 is pmSyn1-EBFP-Cre from Hongkui Zeng’s lab. This AAV has had over 150...

...next experiment. Nucleic acid based machines Recombinase-based State Machines Enable Order-dependent Logic...inputs are received by a cell. These so-called recombinase-based state machines could potentially be used...development. Read this post to learn more about recombinase-based state machines and how they function. Adapting...reminds us that, although fluorescent proteins are incredibly useful, chemical fluorophores can still be used...Dangerous Viruses Contributed by guest blogger Tessa Cressy from the labs of Dr. Elke Mühlberger and Dr. Rachel...CombiGEM and CRISPR for Combinatorial Genetic Screening CombiGEM is a technology that can be used to quickly...barcode small libraries of CRISPR gRNAs to use for screening experiments. Get the full details on how to make...

...can remove the gene through the addition of Cre recombinase (Figure 2). Figure 2: A knockout targeting... can remove a floxed resistance gene using Cre recombinase. If GFP is under control of an endogenous promoter...you'll first remove the resistance marker using Cre recombinase. Since there are 3 loxP sites, recombination...want to create a conditional knockout. To make a conditional knockout, researchers often use the Cre/Lox ...researchers usually remove it after selection using the Cre/Lox recombination system. After “floxing” the resistance...so that it can be removed post-selection through Cre-mediated recombination.   Knockout/Knock-in...hence knock out the gene) through a second round of Cre recombination.  The future of knockout plasmids Although...