Methods for producing a maize plant containing in its genetic material one or more traits introgressed into 2ZZSB1121 through backcross conversion and/or transformation, and to the . [26][27][28] Genetic location of the floxed sequence affects recombination efficiency as well probably by influencing the availability of DNA by Cre recombinase. Answer: a. It is a member of a large family of tyrosine recombinases that includes several other familiar recombinases, such as Flp and Int. Systems, Research PubMedPMID:6276558. 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. Free access to premium services like Tuneln, Mubi and more. NIH Blueprint for Neuroscience Research has created several hundreds of Cre driver mouse lines which are currently used by the worldwide neuroscience community. Each site is 13 bp long, and an 8 bp . Learn about the latest plasmid technologies and research tools. 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). 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. (Figure 1), recombination between [19], Cre recombinase can be synthesized by the corresponding gene under the direction of cell-specific promoters, including promoters under the control of doxycycline. An official website of the United States government. To generate mice that are heterozygous for aloxP-flanked allele and hemizygous/heterozygous for thecretransgene, mate a homozygousloxP-flanked mouse of interest to a cre transgenic mouse strain (see Figure 1 below). mutant/transgenic mice. This reaction links the recombinase protein to the DNA via a phospho-tyrosine linkage. You can search the table for the promoter, fusion, or expression system of choice. lox. By accepting, you agree to the updated privacy policy. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). [26], Holliday junctions and homologous recombination, Implementation of multiple loxP site pairs. The Cre-lox system is a technology that can be used to induce site-specific recombination events. The Cre-Lox system is very similar in action and in usage to the FLP-FRT recombination system. One potential limitation of using loxP sites is the inability to tightly control which loxP sites recombine if more than two are present; intramolecular events happen with greater frequency than intermolecular events, but any two sites can potentially recombine. The following text is excerpted from Siegel et al. 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 3OH end is elongated (that is, bases are added) with the help of DNA Polymerase. recombinase, and loxP sites. Read Addgene's Blog on Cre alternatives from Gerald Rubin's lab. Siegel04.. Bacteriophage P1 uses a site-specific recombination system that is responsible for partitioning newly synthesized genomic copies during replication Abremski, Hoess.This system is composed of a 38-kD phage-encoded Cre recombinase that mediates symmetrical recombination between two 34-bp loxP sites Abremski, which are recreated after . It encodes a site-specific recombinase (q.v.) The resulting mice with both the cre gene and the loxP -flanked gene. The system consists of two components derived from the P1 bacteriophage: the Cre recombinase and a loxP recognition site. We've encountered a problem, please try again. Intro Conditional gene expression using the Cre Lox FLEx vector switch! The Cre-lox system is used as a genetic tool to control site specific recombination events in genomic DNA. 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. During genetic recombination, a Holliday junction is formed between the two strands of DNA and a double-stranded break in a DNA molecule leaves a 3OH end exposed. Engineered lox variants which differ on the spacer region tend to have varied but generally lower recombination efficiency compared to wildtype loxP, presumably through affecting the formation and resolution of recombination intermediate.[25]. Can be used for injection into fertilized zygotes, Cre activates gene of interest by removing neoR and stop sequence; used for making transgenic mice. Heteromeric . To rule out the possibility that the gene recombination efficacy was insufficient for CK19;PTEN / mice to develop tumors, we administered the same dose of TAM for 3 consecutive days by oral gavage, but no ampullary tumors were observed after 90 days (n = 4). Simply put, the Cre/loxsystem rocks. The Cre enzyme leads to the recombination between lox sites removing the marker gene along with one lox site. The .gov means its official. 2007;(178):29-48. doi: 10.1007/978-3-540-35109-2_2. OurTechnical Information Services (TIS)is receiving more and more requests for help in designing breeding schemes for producing Cre/loxmice. The RecA protein is very important for DNA repair; therefore, it is recA dependent recombination. Here is Part 2!). Common construct types include: Note: Through a partnership with genOway, Addgene is able to distribute materials containing FLEx technology. 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. Expression of Cre recombinase by AtRps5apro or AtDD45pro enabled Cre/lox-mediated recombination at an early embryonic developmental stage upon crossing, activating transgenes in the hybrid cowpea and tobacco. The Cre recombinase is an enzyme that is required for 2001 Summer;5(2):81-4. doi: 10.1089/109153601300177600. . In addition, sequence excision can happen in trans instead of a in cis cassette exchange event. If the two daughter plasmids get interlinked one of the daughter cells of the host will lose the plasmid. Again, only one generation of breeding is required. The Lox or loxP sites (locus of X over P1) are the palindromic sites recognized by cre-recombinase. Special features include: Use the search box below to find the perfect Cre expression construct for your experiment. As Cre/ loxP . When researchers started creating Cre recombinase mouse strains to generate tissue specific knockout mice, they needed an easy way to confirm that Cre recombinase was active only in certain tissues. Using different lox site variants allows Cre to catalyze more than one specific recombination event in a single system. 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. Cre Recombinase Cyclization recombinase (Cre) is one of the tyrosine site-specific recombinases, which is known to catalyse the site specific recombination event between two DNA recognition sites (LoxP sites). Subsequent cleavage and rejoining cause DNA strands to exchange their segments. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. pLH1497 ASPNIDRAFT_186610:: lox P-hph-lox P, h y g R, ppt R, kan R This study pLH1498 ASPNIDRAFT_131173 :: lox P- hph - lox P, h y g R , ppt R , kan R This study Content courtesy of Springer . How do I place an order? The Cre-Lox recombination system is derived from P1 Bacteriophages and uses a specific recombinase protein (Cre) that targets a unique DNA sequence (loxP) for splicing. This 3 OH group is highly unstable, and the strand on which it is present must find its complement. A number of conserved site-specific recombination systems have been described in both prokaryotic and eukaryotic organisms. 1992. The schematic below shows the three types of rearrangements: inversion, deletion and translocation. Both 13bp repeat sequences on a single loxP site are recognized and bound by aCre protein, forming a dimer. Altered directionality in the Cre-LoxP site-specific recombination pathway. Clipping is a handy way to collect important slides you want to go back to later. Marth and Rajewsky reported in 1994 that Cre-lox recombination could be used for conditional gene targeting. In general, these systems use one or more proteins and act on unique asymmetric DNA sequences. 14.1k views Gene knockout samuel kwatia 52.1k views Knock out mice Priya Nanda LoxP (locus of X(cross)-over in P1) sites are 34-base-pair long recognition sequences consisting of two 13-bp long palindromic repeats separated by an 8-bp long asymmetric core spacer sequence. The integrase class of enzymes catalyse this particular reaction. Cre (Cre recombinase) Cre-Lox recombination is commonly used to circumvent embryonic lethality caused by systemic inactivation of many genes. Since removal of the stop cassette is permanent, the reporter genes are expressed in all the progeny produced by the initial cells where the Cre was once activated. The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. Plasmids. This recombinase comes from the P1 bacteriophage. Cre reporter strains can also be used to generate a source of labeled tissues/organs, which allow researchers to track cells in transplant or adoptive transfer experiments. Thus, the orientation, and location of the loxP sites are important determinants of This patent application was filed with the USPTO on Wednesday, February 15, 2012 Using an in vivo phagemid system to identify non-compatible loxP sequences. sites, either on -its 8bp asymmetric core or 13-bp palindromic arms. Enjoy access to millions of ebooks, audiobooks, magazines, and more from Scribd. In part 1 of this mini-series, I introduced the concept and applications of Cre-loxP. To do so, one mouse with the Cre recombinase (the Cre This breeding scheme may not be the most efficient one for producing all Cre/loxstrains. In Cre-DIO system, there are two pairs of lox sites (lox site 1 and lox site 2) flanking inverted GOI and reporter gene. The P1 phage is a temperate phage that causes either a lysogenic or lytic cycle when it infects a bacterium. A double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated, resulting in the reciprocal crossover event. 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), A novel maize variety designated X03R618CYFR and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. As with any other technology or research tool, it has limitations and pitfalls that need to be considered while planning experiments or interpreting results. These variant lox sites undergo recombination with other sites of the same type, but are not cross-compatible. The Cre/lox system is a well-established research tool, especially in the field of mouse transgenics. You may not be able to create an account or request plasmids through this website until you upgrade your browser. ADVERTISEMENTS: What do I need to know about the customs and importation process for my country? How can I track requests for my plasmids? Thank you for your participation! To facilitate the use of Cre-lox technology, transgenic mice have been constructed that express Cre under a variety of ubiquitous and regulated promoters, and many loxP-containing alleles have also been constructed. Cre, or cyclic recombinase, is one such enzyme. These will be your experimental mice. We archive and distribute high quality plasmids from your colleagues. Site-specific recombination (SSR) involves specific sites for the catalyzing action of special enzymes called recombinases. Cre can invert a region rather than delete it. the loxP sites will delete the gene The Cre-lox system is a technology that can be used to induce site-specific recombination events. cre-lox and cre recombinases in Mouse Genome Informatics (MGI): Module 2 MouseGenomeInformatics 17.7k views Knock out technology (final) Dr Vijayata choudhary 22.2k views Gene knockout GGS Medical College/Baba Farid Univ.of Health Sciences. Figure Recombination involve pairing between complementary strands of the two parental DNAs. 1. Dear Qu, Please read the following: In the field of genetics, Cre-Lox recombination is known as a site-specific recombinase technology, and is widely used to . Activate your 30 day free trialto continue reading. By undergoing Cre recombination, the region floxed by the loxP sites will become inverted, [12] this process is not permanent and can be reversed. instance, if two loxP sites are placed on different chromosomes, a translocation can take The Cre/lox-site-specic recombination system derived from bacteriophage P1 is known to be appropriate for genome modication [23-26], and its mechanism of action has been clearly described [27-29]. All cell types tested in mice have been shown to undergo transgenic Cre recombination. To create a conditional knock-out mouse, one needs the following systems: the Cre-loxP, and Assistant Professor (Ad Hoc) What strain of bacteria does my stab contain? Consistent with functional p53-siRNA expression by these vectors, Cre-mediated recombination resulted in a dramatic reduction of both p53 mRNA and protein levels in pSico-p53-infected cells ( Fig. 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 recombination is a site-specific recombinase technology widely used to carry out deletions, insertions, translocations and inversions in the DNA of cells. Addgene is a nonprofit plasmid repository. Addgene has a wide variety of Cre recombinase plasmids. More novel and sophisticated uses for the Cre/loxsystem are being developed all the time.
IHDR y ] + sRGB pHYs od FIDATx^.{s" ""S 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 . Chai W, Hao W, Liu J, Han Z, Chang S, Cheng L, Sun M, Yan G, Liu Z, Liu Y, Zhang G, Xing L, Chen H, Liu P. JBMR Plus. Federal government websites often end in .gov or .mil. The Saccharomyces cerevisiae FLP-FRT recombination system is another site-directed recombination technology very conceptually similar to Cre-lox, with flippase (Flp) and the short flippase recognition target (FRT) site being analogous to Cre and loxP, respectively. Both AAV and Lenti preps of Cre are available. Here, we present a simple method that combines the use of CRISPR-Cas9 and Cre-LoxP for the Recombinant modified Cre recombinase protein translocates into the cytoplasm and nuc Looks like youve clipped this slide to already. The Cre activity can be induced by applying inducer to initiate the recombination which will lead to self-excision of cre and the marker gene (see below) Marker gene Trait gene loxP loxP Chemical-induced cre gene chemical Trait gene loxP Genomic targeting with a positive-selection lox . eCollection 2023 Jan. Wang N, Lv L, Huang X, Shi M, Dai Y, Wei Y, Xu B, Fu C, Huang H, Shi H, Liu Y, Hu X, Qin D. Front Mol Neurosci. FEBS Lett. The Cre/ lox system is one of the most powerful and versatile tools developed for mouse genetics. In the absence of tamoxifen, CreER will result in the shuttling of the mutated recombinase into the cytoplasm. Cells alter their phenotype in response to numerous environmental stimuli and can lose the expression of genes typically used to mark their identity, making it difficult to research the contribution of certain cell types to disease. The SlideShare family just got bigger. Cell Culture Media, Introduction to the Cre-Lox Recombination System, AAALAC-accredited and OLAW assured animal centers, Specific-pathogen-free (SPF) animal health status exceeds requirements of most facilities, Top-tier customer support: free consultations and quotes. At a DNA level many types of manipulations can be carried out. The protein will stay in this location in its inactivated state until tamoxifen is given. Methods for producing a maize plant comprise crossing maize variety 2ZZSB1121 with another maize plant are provided. Dre-Rox system. Because the controls will not have Cre recombinase, any phenotypic differences between them and the Cre/lox mice should be due to the deleted gene in the Cre/lox mice. First, the nucleotide sequence identity in the spacer region of lox site. More specifically, it relates to methods and compositions for the introduction of DNA using circular molecules that are not able to replicate outside a host cell. Since both the transposon and the Cre- lox recombineering systems were applied to diverse hosts, CRAGE enabled insertion of large biosynthetic gene clusters into 25 diverse gammaproteobacterial species. Maize hybrid X13C737 is an invention by Andrew Jon Ross, Lisbon IA UNITED STATES. A soybean cultivar designated 4826008 is disclosed. The Cre/lox system can be used to turn off a transgene. The Cre-lox system serves several functions in the phage: it circularizes the phage DNA into a plasmid, separates interlinked plasmid rings so they are passed to both daughter bacteria equally and may help maintain copy numbers through an alternative means of replication. The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. A novel maize variety designated 2ZZSB1121 and seed, plants and plant parts thereof are provided. government site. Inducible Cre can be leaky. [20]. We've updated our privacy policy. Looks like youve clipped this slide to already. Site-specific recombination is, thus, the enzyme-mediated cleavage and ligation of two defined deoxynucleotide sequences. For many years, it was thought that when the Holliday junction intermediate was formed, the branch point of the junction (where the strands cross over) would be located at the first cleavage site. It was unclear whether only one allele in each T-cell or 50% of T cells had 100% deletion in both alleles. It gives mouse researchers sophisticated control over the location and timing of gene expression. The Flp-FRT system and Cre-loxP induce gene recombination in a similar way. Digvijay Autonomous P. G. College 2001 Jun 15;499(1-2):147-53. doi: 10.1016/s0014-5793(01)02541-8. hbspt.cta._relativeUrls=true;hbspt.cta.load(306096, 'be59770e-eb9c-43af-8b8e-a9e2262f9e74', {"useNewLoader":"true","region":"na1"}); Cre recombinase, originally named because it causes recombination (although later referred to as the cyclization recombinase), is a 38 kDa protein responsible for intra- and inter-molecular recombination at the loxP recognition sites. PubMed, CRISPR Expression Systems and Delivery Methods. Careers. 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. This prevents non-specific actions of Cre recombinase. The loxP site (34bp) has a central These sites, known as loxP sequences, contain specific binding sites for Cre that surround a directional core sequence where recombination can occur. 3. Sauer, B. and Henderson, N. 1988. Another factor is the length of DNA between the lox pair. 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.). Translocation: If the sites are on separate DNA molecules, a translocation event is generated at the loxP sites. Faust TE, Feinberg PA, O Connor C, Kawaguchi R, Chan A, Strasburger H, Masuda T, Amann L, Knobeloch KP, Prinz M, Schaefer A, Schafer DP. Plasmids 101, Front Immunol. Cre is a 38 kDa recombinase protein from bacteriophage P1 that catalyzes recombination between LoxP sites. NNNTANNN", where "N" indicates which bases may vary from the canonical sequence, Sternberg, N. and Hamilton, D. 1981. The pairing of opposite strands is what constitutes the crossing-over or Recombination event, which is common to all living organisms, since the genetic material on one strand of one duplex has paired with one strand of another duplex, and has been elongated by DNA polymerase. Notice that only one generation of breeding is required (Hooray!). recombination; recombination between two loxP sites can induce a deletion. cre-lox and cre recombinases in Mouse Genome Informatics (MGI): Module 2. The two loxP sites then align in a parallel orientation, allowing the four Cre proteins to form a tetramer. When there are In this context we created four target site lines for RMCE and evaluated their fitness costs. found in the yeast and based on the yeast two-micron plasmid. (RBE); the two RBE are 13bp Explanation: The Cre recombinase protein of bacteriophage P1 mediates site-specific recombination at a 34 bp sequence, loxP. lox. Diversity and Selection of Shell of the Hermit Crab of Mandvi, Kachchh Coast, No public clipboards found for this slide, Enjoy access to millions of presentations, documents, ebooks, audiobooks, magazines, and more. Cleavage on the other strand also causes a phospho-tyrosine bond between DNA and the enzyme. 2022 Dec 14;15:1043018. doi: 10.3389/fnmol.2022.1043018. Applied Biological Materials - abm 40.7K subscribers Subscribe 1.4K 102K views 3 years ago The FLEx vector or "flip. An important application of the Cre-lox system is excision of selectable markers in gene replacement. See, for example, Lyznik et al., Site-Specific Recombination for Genetic Engineering in Plants, Plant Cell Rep, 21:925-932 (2003) and WO 99/25821, which are hereby incorporated by reference. Well, it enables them to generate tissue-specific and inducible knockouts and thereby have exquisite control over the location and timing of gene expression important stuff when deleting a certain gene everywhere or during development leads to an embryonic lethal phenotype. mouse), and another with the loxP sites (the floxed mouse) are created, and crossed to. 2. Before We've updated our privacy policy. A base pair (bp) is a unit consisting of two nucleobases bound to each other by . called the FLP (pronounced 'flip') protein. 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. Visualizing Cathepsin K-Cre Expression at the Single-Cell Level with GFP Reporters. Site-specific recombination is also an important process that viruses, such as bacteriophages, adopt to integrate their genetic material into the infected host. One of my colleagues, Andy, has written an awesome blog post on howCre recombinasecan produce a phenotype on its own. The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. Do you have any tips or tricks, or clever cre-dependent techniques? . 11. Cre (shown at left) is a bacteriophage P1 member of the integrase family, catalyzing site-specific recombination between two, 34-base pair lox DNA sequences. sites contain nucleotide variations in the left or right arms, respectively named LE and RE. Approximately 50% of the offspring will be heterozygous for theloxPallele and hemizygous/heterozygous for thecretransgene. "Site-specific recombination of DNA in eukaryotic cells", "Functional expression of the cre-lox site-specific recombination system in the yeast Saccharomyces cerevisiae", "Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1", "Tissue- and site-specific DNA recombination in transgenic mice", "T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination", "Cre-Lox Neurogenetics: 20 Years of Versatile Applications in Brain Research and Counting", "Subregion- and cell type-restricted gene knockout in mouse brain", "The essential role of hippocampal CA1 NMDA receptor-dependent synaptic plasticity in spatial memory", http://www.neuroscienceblueprint.nih.gov/factSheet/CreDriver.htm, "Skeletal and CNS defects in Presenilin-1-deficient mice", "Deficient neurogenesis in forebrain-specific presenilin-1 knockout mice is associated with reduced clearance of hippocampal memory traces", "Yeast two-hybrid interaction partner screening through in vivo Cre-mediated Binary Interaction Tag generation", "Cyclization recombinase [Escherichia coli] - Protein - NCBI", "Targeted integration of DNA using mutant lox sites in embryonic stem cells", "A high-throughput screen identifying sequence and promiscuity characteristics of the loxP spacer region in Cre-mediated recombination", "A genetic strategy for stochastic gene activation with regulated sparseness (STARS)", "Engineering mouse chromosomes with Cre-loxP: range, efficiency, and somatic applications", "Non-parallel recombination limits Cre-LoxP-based reporters as precise indicators of conditional genetic manipulation", "Genetically engineered mouse models in cancer research", "Spontaneous recombinase activity of Cre-ERT2 in vivo", "Epigenetic Regulation of Vascular Smooth Muscle Cells by Histone H3 Lysine 9 Dimethylation Attenuates Target Gene-Induction by Inflammatory Signaling", "Extensive Proliferation of a Subset of Differentiated, yet Plastic, Medial Vascular Smooth Muscle Cells Contributes to Neointimal Formation in Mouse Injury and Atherosclerosis Models", "Previously differentiated medial vascular smooth muscle cells contribute to neointima formation following vascular injury", "KLF4-dependent phenotypic modulation of smooth muscle cells has a key role in atherosclerotic plaque pathogenesis", "Lineage tracing of cells involved in atherosclerosis", "Disease-relevant transcriptional signatures identified in individual smooth muscle cells from healthy mouse vessels", Introduction to Cre-lox technology by the "Jackson Laboratory", https://en.wikipedia.org/w/index.php?title=Cre-Lox_recombination&oldid=1135942402, This page was last edited on 27 January 2023, at 20:38. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. 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. Activate your 30 day free trialto unlock unlimited reading. Why? Comparative mitochondrial zygomycetes: bacterial-like Rnase P RNAs, mobile el No public clipboards found for this slide, Enjoy access to millions of presentations, documents, ebooks, audiobooks, magazines, and more. 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. [29] This is done through the fusion of a mutated ligand binding domain of the estrogen receptor to the Cre recombinase, resulting in Cre becoming specifically activated by tamoxifen. Cre-ERT2 - Tamoxifen inducible; Gateway entry vector, Cre expressed at low levels to reduce toxicity; See PI page for other recombinases that are less toxic in Drosophila, Cre-EGFP fusion; Tet inducible - rrTA expression driven by mouse Nkx cardiac enhancer and promoter fragment, N-terminal component of the Co-InCre system, C-terminal component of the Co-InCre system, Cre-ERT2; Targeting vector for Nanog locus, Cre, KASH-tagged EGFP, and sgRNA expression, DHFR-destabilized Cre; targeting vector for Rasgrf2, Cre-ERT2 - Tamoxifen inducible; Targeting vector, Cre fused to supernegatively charged GFP variant, Cre fused to the human Estrogen Binding Domain (EBD), Cre with a 25 nucleotide extracellular vesicle targeting sequence and CFP, mCherry and Cre expression in newborn neurons, GFP and Cre expression in newborn neurons, Cre recombinase dependent on GFP (CRE-DOG), iCre with MCS for inserting promoter, WPRE, CreER expression and tetracyclin-dependent transgene/shRNA expression, mCherry and Cre; expressed in excitatory neurons, For in vitro transcription of Cre or to recombine into BAC, TFP and Cre-ERT2 - Tamoxifen inducible.