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Bacterial genome editing by coupling Cre- lox and CRISPR-Cas9 - PLOS Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. Researchers have since reported more efficient Cre-Lox conditional gene mutagenesis in the developing T cells by the Marth laboratory in 1995. Gu, H., Zou, Y.R., and Rajewsky, K. 1993. Cre/ lox -mediated recombination in Arabidopsis - Academia.edu The obvious difference between the two systems is that the recombinase (Cre and Flp) has different optimal reaction temperatures. About 25% of the offspring from this mating will be homozygous for theloxPflanked allele but will have nocretransgene. The caveat, however, is that the recombination reaction can happen backwards, rendering cassette exchange inefficient. Scientists have developed ways to tightly control Cre expression and to optimize Cre expression once it is induced. Cre-Lox recombination is a site-specific recombinase technology, used to carry out deletions, insertions, translocations and inversions at specific sites in the DNA of cells. Cre-lox Systems - Rockefeller University Below, I would to focus on two additional issues: using Cre/lox mice to turn on or off transgenes, and using Cre reporter strains. & Engineering, Model Cre-lox System | Cyagen Korea The Cre-lox recombination system prevents these situations by unlinking the rings of DNA by carrying out two recombination events (linked rings -> single fused ring -> two unlinked rings). [13] Mechanism of translocation [ edit] official website and that any information you provide is encrypted a) The chromosomal copy of the target gene replaces the target gene flanked by loxP sites. Cre-lox in vivo/vitro . Please note: Your browser does not support the features used on Addgene's website. Let us know in the comments! Have questions about your order, deposit, or a plasmid? RuvC is just one of these Resolvases that have been isolated in bacteria and yeast. Research mouse models that incorporate the Cre/loxsystem typically involve generating double,triple, quadruple, or evenn-tuple(is that really a word?) Cre-lox ? These targeted DNA changes are useful in cell lineage tracing and when mutants are lethal if expressed globally. . * One unit is defined as the amount of enzyme necessary to produce maximal site-specific recombination of 0.25 g pLox2+ control DNA in 30 minutes at 37C in a total reaction volume of 50 l. A novel maize variety designated X00R808 and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. Cre Lox Recombination | PDF | Life Sciences | Molecular Biology This conserves the energy of the phosphodiester bond, allowing the reaction to be reversed without the involvement of a high-energy cofactor. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X03R618CYFR through backcrossing . In this context we created four target site lines for RMCE and evaluated their fitness costs. The mechanism of Cre recombination requires two lox sites be brought together by looping the intervening DNA. Considering users' convenience to optimize the amount of blue light needed for single genomic Cre-lox recombination, we infected PA-Cre 3.0 to MEFs isolated from Ai14: Floxed-tdTomato . Cre-Lox: A Tool for Removal of Marker Genes to Make GM Foods Safe Read Addgene's Blog on Cre alternatives from Gerald Rubin's lab. The versatile properties of Cre recombinase make it ideal for use in many genetic engineering strategies. Methods for producing a maize plant containing in its genetic material one or more traits introgressed into X00R808 through backcrossing or genetic . Effective production of kojic acid in engineered Aspergillus niger mutant/transgenic mice. The enzyme (38kDa) is a member of the integrase family of site specific recombinase and it is known to catalyse the site specific recombination event between two DNA recognition sites (). Can be used for injection into fertilized zygotes, Cre activates gene of interest by removing neoR and stop sequence; used for making transgenic mice. Unable to load your collection due to an error, Unable to load your delegates due to an error. ADVERTISEMENTS: Phages like the lambda phage use their site specific recombinases to integrate their DNA into the host genome during lysogeny. cre lox recombination ppt FEBS Lett. Activate your 30 day free trialto unlock unlimited reading. 2023 Jan 9:2023.01.09.523268. doi: 10.1101/2023.01.09.523268. Both AAV and Lenti preps of Cre are available. Cre-lox-regulated conditional RNA interference from transgenes Now, one point of crossover has occurred, which is what is called a Holliday Intermediate. Cre-lox , , knockout , . Recombination: Definition, Mechanism and Types | Microbiology Cre-Lox recombination involves the targeting of a specific sequence of DNA and splicing it with the help of an enzyme called Cre recombinase. 2022 Dec 21;7(1):e10706. For So, in this post, Im outlining a typical breeding scheme that I hope will help demystify the process of using the Cre/loxsystem to generate tissue-specific or inducible knockout mice. Site-specific recombination (SSR) involves specific sites for the catalyzing action of special enzymes called recombinases. Studies have found that Cre recombinase is the best The temperature is 37 oC and Flp recombinase is 30. PubMed, CRISPR Expression Systems and Delivery Methods. The Cre-lox system, derived from P1 bacteriophage, is a potent and specific system for controlling gene expression. Introduction Cre-lox recombination Cre-lox system- Cre recombinase , loxP site FLP-FRT recombination FLP-FRT system- FLP recombinase , FRT site Mechanism of Cre-lox and FLP-FRT recombination Binding Synapsis , cleavage and strand exchange Three type of arrangement Inversion Translocation/ Insersion Deletion Application of Cre-lox and FLP-FRT recombination Disadvantage of FLP-FRT Advantage and disadvantage of Cre-lox Conclusion References. In the absence of tamoxifen, CreER will result in the shuttling of the mutated recombinase into the cytoplasm. doi: 10.1002/jbm4.10706. OMP targeting vector with Cre FNF (neo-selectable marker flanked by FRT sites), M71 targeting vector with IRES CreFNF (neo-selectable marker flanked by FRT sites), Cre, Puro resistance and miRNA expression. 2007;(178):29-48. doi: 10.1007/978-3-540-35109-2_2. (In a future post, I will tackle the breeding schemes for using the system to turn transgenes on or off. 5 Phosphate ends are usually the substrates for this reaction, thus extended 3 regions remain. The technology called Cre-lox recombination relies on the use of an enzyme called Cre recombinase which can bind, cut and recombine DNA at specific sites that are inserted in pairs in the DNA. These strains are designed to express a visible marker, such as green fluorescent protein (GFP) or LacZ, only after Cre recombinase excises a loxP-flanked stop sequence (Figure 7 and 8.). Upon Cre driven recombination, the stop cassette is excised, allowing reporter genes to express specifically in cells in which the Cre expression is being driven by the cell-specific marker promoter. If you run into any problems registering, depositing, or ordering please contact us at [emailprotected] Cre/loxP Recombination System: Applications, Best Practices - Taconic MeSH The Cre-lox system is a technology that can be used to induce site-specific recombination events. This reaction links the recombinase protein to the DNA via a phospho-tyrosine linkage. The Cre-Lox recombination is quite a simple tool to use and manipulate, thus it has been widely used in science since the discovery of Cre more thant 30 years ago (1). 4. US Patent for Maize hybrid X00R808 Patent (Patent # 11,589,542 issued Answer: a. Here is the most efficient breeding scheme for generating Cre/loxtissue-specific or inducible knockouts. You will have to adapt it to the genotypes you want to produce and the genetic backgrounds and characteristics of theloxP-flanked andcrestrains you use. Models thus obtained are more likely to mimic the physiological situation. Removal of selectable markers from the genome by Cre-lox recombination is an elegant and efficient way to circumvent this problem and is therefore widely used in plants, mouse cell lines, yeast, etc.[1]. Both 13bp repeat sequences on a single loxP site are recognized and bound by aCre protein, forming a dimer. The site includes an asymmetric 8 bp sequence, variable except for the middle two bases, in between two sets of symmetric, 13 bp sequences. This plasmid is an autonomously replicating, circular plasmid of 6,318 base pairs, which exists in many copies in most strains of S. cerevisiae (q.v.). This patent application was filed with the USPTO on Wednesday, February 15, 2012 The Lox or loxP sites (locus of X over P1) are the palindromic sites recognized by cre-recombinase. Nov 2, 2015. We've updated our privacy policy. Deletion: If the sites face in the same direction, the sequence between the loxP sites is excised as a circular piece of DNA (and is not maintained). This dimer then binds to a dimer on another lox site to form a tetramer. While the use of the cre-loxP system is becoming increasingly widespread, in particular because of growing availability of conditional mouse mutants, many considerations need to be taken into account when utilizing the cre-loxP system. Application of Cre-loxP system to the urinary tract and cancer gene therapy. [38], A classical strategy for generating gene deletion variants is based on double cross-integration of non-replicating vectors into the genome. Do you have suggestions for other plasmids that should be added to this list? Among these are loxN (GtATACcT), lox2272 (GgATACtT), and lox511 (GtATACAT). sites contain nucleotide variations in the left or right arms, respectively named LE and RE. 2017 by John Wiley & Sons, Inc. Keywords: 1. How can I be notified when a plasmid from a specific lab or paper is available? Conversely, pSicoR-p53 generated a p53 knock-down that was reversed upon Ad-Cre infection ( Fig. The Cre- lox recombination system is derived from the bacteriophage P1. In more recent years, however, this view has been challenged, and most of the current models for Int, Xer, and Flp recombination involve only limited branch migration (13 base pairs of the Holliday intermediate), coupled to an isomerisation event that is responsible for switching the strand cleavage specificity. PDF A Novel Cre/lox-Based Genetic Tool for Repeated, Targeted and Bibliography : If the two loxP sites are in the same orientation, the floxed sequence (sequence flanked by two loxP sites) is excised; however if the two loxP sites are in the opposite orientation, the floxed sequence is inverted. The invention relates to the seeds of soybean cultivar 03070412, to the plants of soybean cultivar 03070412, to the plant parts of soybean cultivar 03070412, and to methods for producing progeny of soybean cultivar 03070412. You can search the table for the promoter, fusion, or expression system of choice. Instant access to millions of ebooks, audiobooks, magazines, podcasts and more. By whitelisting SlideShare on your ad-blocker, you are supporting our community of content creators. . PubMed CentralPMCID:PMC49604. An official website of the United States government. This model provided convenient explanation for the strict requirement for homology between recombining sites, since branch migration would stall at a mismatch and would not allow the second strand exchange to occur. Cre-Lox recombination - Unionpedia, the concept map Colorful Retro Vintage Illustration Animated Medical Technology Education Sci Quantized space time and Einstein's special theory of relativity.pdf, Doodle Astronomy Lesson XL by Slidesgo.pptx. Department of Biotechnology The present invention relates generally to transgenic plants. What is an MTA/Who is authorized to sign? Cre/lox switchingexpression system Currently, mostwidely used site-specific DNA recombination system system[18, 19]. Empty backbones to regulate your gene of interest: In this subtype of loxP plasmids, reporter genes indicate which cells have undergone Cre recombination. Cre activates gene of interest, removes Neo cassette. Shortly thereafter, researchers in the laboratory of Prof. Klaus Rajewsky reported the production of pluripotent embryonic stem cells bearing a targeted loxP-flanked (floxed) DNA polymerase gene. The resulting mice with both the cre gene and the loxP -flanked gene. During site-specific DNA recombination, which brings about genetic rearrangement in processes such as viral integration and excision and chromosomal segregation, these recombinase enzymes recognize specific DNA sequences and catalyse the reciprocal exchange of DNA strands between these sites. Targeting Your Dna With the Cre/Lox System | Scq Non-parallel recombination is especially problematic in a fate mapping scenario where one recombination event is designed to manipulate the gene under study and the other recombination event is necessary for activating a reporter gene (usually encoding a fluorescent protein) for cell lineage tracing. FLP-FRT & Cre-lox Recombination The double stranded DNA is cut at both loxP sites by the Cre protein. The Cre enzyme and the original Lox site called the LoxP sequence are derived from bacteriophage P1. Migration of the branch point to the second cleavage site would then somehow trigger the second half of the pathway. If you are looking for the right Cre-expressing strain for your experiments, you can search for strains at the JAX Cre Repository or through the Cre Portal at MGI. Cre-Lox recombination - Wikipedia Do not sell or share my personal information, 1. If loxP sites are on different chromosomes it is possible for translocation events to be catalysed by Cre induced recombination. A key advantage of the system is that Cre acts independently of any other accessory proteins or co-factors, thus allowing for broad applications in a variety of experiments. Is LysM-Cre a good candidate Cre for knocking out Atg5 gene in mice. For a more thorough introduction, check out Addgenes Cre-lox blog post. Methods for producing a maize plant by crossing hybrid maize variety X00R808 with another maize plant are disclosed. A system of site-specific recombination (q.v.) [40]. In previous posts for our Plasmids 101 series, we examined a number of important plasmid elements promoters, origins of replication, protein tags, and antibiotic resistance markers(just to name a few). It is implemented both in eukaryotic and prokaryotic systems. For example, mating the transgenic strain in Figure 1 to a strain that expresses Cre recombinase in mammary tissue produces double transgenic offspring that express theKras oncogene only in the mammary glands (Figure 5). The C domain is similar in structure to the domain in the Integrase family of enzymes isolated from lambda phage. conditional genetics; cre; cre-loxP; gene editing; loxP; recombination; site directed mutagenesis. The Cre/lox system is one of the most powerful and versatile tools developed for mouse genetics. [30] Importantly, sometimes fluorescent reporters can be activated in the absence of tamoxifen, due to leakage of a few Cre recombinase molecules into the nucleus which, in combination with very sensitive reporters, results in unintended cell labelling. Cre-Lox recombination - Columbia University Postdoc Society Blog First, let us focus on the Cre-loxP system which requires a Cre Cre can invert a region rather than delete it. recombination is reversible, strategies for irreversible integration rely on mutated . Conditional knockout mice are usually generated using Cre-LoxP, Dre-Rox or Flp-Frt recombination systems for permanent gene alterations in specific tissues. Before Depending on the Depending on your experimental parameters, you can use these as controls (see Figure 3A below). While Cre-lox recombination is a very useful tool, Cre expression can be toxic in some systems, notably Drosophila. 1. found in the yeast and based on the yeast two-micron plasmid. Protein-protein interactions drive and direct strand exchange. Cre-lox Systems Cre-lox (floxed) systems for gene deletion ( click here for non-floxed and earlier floxed tagging vectors) General Comments The baseline reference for this system is CRE recombinase-based positive-negative selection systems for genetic manipulation in Trypanosoma brucei (2008) Scahill MD, Pastar I & Cross GAM. Based on these Cre-lox recombination principles, scientists have developed constructs to activate/inactivate genes when Cre is present. US Patent for Soybean cultivar 4826008 Patent (Patent # 11,589,543 The 3OH end is elongated (that is, bases are added) with the help of DNA Polymerase. P1 phage DNA on the other hand, exists as a plasmid in the host. Does Addgene accept orders by fax, phone or email? It was unclear whether only one allele in each T-cell or 50% of T cells had 100% deletion in both alleles. In the lysogenic cycle the phage genome replicates with the rest of the bacterial genome and is transmitted to daughter cells at each subsequent cell division. Thank you for your participation! In particular, the clear demonstration of its usefulness in precisely defining the complex relationship between specific cells/circuits and behaviors for brain research,[12] has promoted the NIH to initiate the NIH Blueprint for Neuroscience Research Cre-driver mouse projects in early 2000. It is one of the most powerful tools in the mouse geneticists toolbox. Tissue and sitespecific recombination in transgenic mice. Activate your 30 day free trialto unlock unlimited reading. Lecture 11 - Lectures For UG-5 - studyres.com The protein will stay in this location in its inactivated state until tamoxifen is given. Two factors have been shown to affect the efficiency of Cre's excision on the lox pair. These lox variants only proceed recombination with the same type of lox sites, with no ability to interact with the other types. It appears that you have an ad-blocker running. 11. Cre-lox recombination, discovered in the 1980s, is one of the most important ways to spatially and temporally control gene expression, especially in in vivo models, and new Cre-lox based technologies are still being developed today. The Cre/lox system can be used to turn off a transgene. Cre-Lox recombination is a site-specific recombinase technology widely used to carry out deletions, insertions, translocations and inversions in the DNA of cells. must be introduced artificially. The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. This event can have three general outcomes based on the location and orientation of the loxP sites: Inversion: If the loxP sites are on the same DNA strand and are in opposite orientations, recombination results in an inversion and the region of DNAbetween the loxP sites is reversed. It is also proposed that rolling circle replication followed by recombination will allow the plasmid to increase its copy number when certain regulators (repA) are limiting. [13][14] To date, NIH Blueprint for Neuroscience Research Cre projects have created several hundreds of Cre driver mouse lines which are currently used by the worldwide neuroscience community. The SlideShare family just got bigger. Matsumura T, Watanabe T, Totsugawa T, et al. sites, either on -its 8bp asymmetric core or 13-bp palindromic arms. Cre recombinase was used to mediate recom- Introduction bination between a chromosomally introduced loxP se- quence in Arabidopsis thaliana (35S-lox-cre) and trans- Site-specific recombination systems from prokaryotes ferred DNA (T-DNA) originating from Agrobacterium and yeast, such as the bacteriophage P1 Cre/lox system, tumefaciens (plox-npt), The Cre recombinase is an enzyme that is required for [3][4][5] Subsequently, researchers in the laboratory of Dr. Jamey Marth demonstrated that Cre-Lox recombination could be used to delete loxP-flanked chromosomal DNA sequences at high efficiency in specific developing T-cells of transgenic animals, with the authors proposing that this approach could be used to define endogenous gene function in specific cell types, indelibly mark progenitors in cell fate determination studies, induce specific chromosomal rearrangements for biological and disease modeling, and determine the roles of early genetic lesions in disease (and phenotype) maintenance.[6]. Addgene has a wide variety of Cre recombinase plasmids. Cre activates gene of interest, using 4 mutated lox sites, Cre-loxP recombination is used to create a stochastic choice of expression among fluorescent proteins, resulting in the indelible marking of mouse neurons with multiple distinct colors, Multibow uses a binary code in which reporters are initially OFF and then probabilistically ON or OFF following Cre recombination to maximize label diversity, Rosa26 targeting vector, Cre dependent tdtomato expression. This recombinase comes from the P1 bacteriophage. PDF Cre/lox-mediated chromosomal integration of biosynthetic gene clusters Cleavage on the other strand also causes a phospho-tyrosine bond between DNA and the enzyme. loxP (locus of X-over P1) is a site on the bacteriophage P1 consisting of 34 bp. This tool is suitable for deleting antibiotic resistance genes, but above all it allows conditional knockouts that can be induced at specific times in the cell type of choice. The fluorescent reporter cassette will contain a promoter to permit high expression of the fluorescent transgene reporter (e.g. place. GENE KNOCKOUT USING THE CRE-LOXP SYSTEM For a few examples of such strains, check out 008179 and 006481. Many other proteins apart from the recombinase are involved in regulating the reaction. Clipping is a handy way to collect important slides you want to go back to later. Instant access to millions of ebooks, audiobooks, magazines, podcasts and more. Disclaimer. This site needs JavaScript to work properly. [28] Failure to activate both recombination events simultaneously confounds the interpretation of cell fate mapping results. Lox sites are directional and the two sites joined by the tetramer are parallel in orientation. The following text is excerpted from Siegel et al. KAUSHAL KUMAR SAHU AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR, AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR, AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR, AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA, pGEMT-Pax7bait-P2a-GAP43-Cherry-T2a-mERt-Cre-Ert, pCAGGS-mTagBFP2-T2A-sfGFP-GSAx9-iCre-ERT2, pCAG-Synaptophysin-TdTomato-IRES2-OHT-Cre, hsp70l-loxP-mCherry-STOP-loxP-H2B-GFP_cryaa-cerulean, pJFRC172-10XUAS-loxP>-dSTOP-loxP>-myr::GFP, Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientists View, CREM and CMV-STOP-luc cassette, separated by insulator, Cre-EGFP fusion in a promoterless vector with several available restriction sites, tet inducible Cre-GFP fusion, metallothionein MT-I region including the polyadenylation site and several introns, Cre-ERT2 with loxp cassette; Targeting vector for Rosa26 locus, CREM fused to mutant AR ligand binding domain, Cre fused to HIV-TAT - promotes cellular uptake of recombinant Cre, Cre-Myc, Expressed in rod photorecetor cells in the retina. Raj-Nandgaon ( C. G. ). Here is Part 2!). By accepting, you agree to the updated privacy policy. Maize hybrid X13C733 Patent | Application Number: 13396674 300+ TOP MCQs on Cre Lox Excision and Answers 2023 Cre-loxP-Mediated Recombination: General Principles and - PubMed How Does Cre, Flp, Dre Recombinase System Work? I highly recommend that you check out his blog. To verify that Cre recombinase expression does not contribute to a phenotype of interest, I suggest using thecrestrain itself (without anyloxP-flanked alleles) as a control (Figure 3B). See, Heat-inducible reporter with Cre dependent H2B-RFP expression, Switches from DsRed to eGFP expression upon the presence of Cre, Zsgreen is expressed in cells without Cre, mCherry is expressed with Cre, Double fluorescent, double selectable cre/loxP reporter, Reporter construct for lineage tracing and mosaic analysis in zebrafish. Common construct types include: Note: Through a partnership with genOway, Addgene is able to distribute materials containing FLEx technology. Activate your 30 day free trialto continue reading. 2022 Aug 26;13:964496. doi: 10.3389/fimmu.2022.964496. This system has allowed researchers to manipulate a variety of genetically modified organisms to control gene expression, delete undesired DNA sequences and modify chromosome architecture. It addresses the limitations of cre-loxP technology and related considerations for experimental design, and it discusses alternative strategies for site-specific genetic recombination and integration. Furthermore, recombination systems such as Cre-lox are widely used, mostly in eukaryotes. Cre-lox, Enjoy access to millions of ebooks, audiobooks, magazines, and more from Scribd. The system consists of two components derived from the P1 bacteriophage: the Cre recombinase and a loxP recognition site. Sternberg, N. and Hamilton, D. 1981. Expressed in chondrocytes, mCherry and Cre-ERT2 - Tamoxifen inducible; Targeting vector for Pax7, Venus and Cre-ERT2 with MCS for inserting promoter, Emerald and Cre-ERT2 with MCS for inserting promoter, TFP and Cre-ERT2 with MCS for inserting promoter, tdTomato and Cre-ERT2 with MCS for inserting promoter, Venus, Cre-ERT2, targeting vector with MCS for homology arms, Emerald, Cre-ERT2, targeting vector with MCS for homology arms, TFP, Cre-ERT2, targeting vector with MCS for homology arms, tdTomato, Cre-ERT2, targeting vector with MCS for homology arms, GAG-Cre fusion; for the production of VLPs loaded with CRE protein, sfGFP-iCre-ERT2 (PAPGSTM N-terminus, unskippable linker) - Tamoxifen inducible, sfGFP-iCre-ERT2 (PAPGSTM N-terminus, GSAx9 linker) - Tamoxifen inducible, iCre-ERT2 (PAPGSTMA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PV N-terminus) - Tamoxifen inducible, iCre-ERT2 (PVV N-terminus) - Tamoxifen inducible, iCre-ERT2 (PVGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PA N-terminus) - Tamoxifen inducible, CreLite - Red light-inducible Cre; N terminus Cre fused to PIF6, CreLite - Red light-inducible Cre; C terminus Cre fused to PhyBdelta, CreLite system components, PhyBdeltaCreC and PIF6CreN, in middle entry vector (Tol2 kit), CreLite; Tol2 destination vector with mTagBFP2, CreLite system components, PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer vector, CreLite system components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector, Cre recombinase split with Vivid photodimers, Cre-ERT2 - Tamoxifen inducible; AAV donor vector, iCre-ERT2 (PAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAAA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAAGSA N-terminus) - Tamoxifen inducible, iCre-ERT2 (PAGSAS N-terminus) - Tamoxifen inducible, N-terminal Cre component split with Vivid photodimers, C-terminal Cre component split with Vivid photodimers, Split Cre fused to nuclear localized wild-type VVD, Retinal ganglion cell-specific expression of Cre, Cre activates shRNA expression, removal of EGFP; See also similar plasmids, Cre turns off shRNA expression, removal of EGFP; See also similar plasmids, Cre activates gene of interest by removing Stop sequences; Puro selection.
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