The Cre- lox recombination system is derived from the bacteriophage P1. Gu, H., Zou, Y.R., and Rajewsky, K. 1993. In addition, sequence excision can happen in trans instead of a in cis cassette exchange event. A high-throughput screen identifying sequence and promiscuity characteristics of the loxP spacer region in Cre-mediated recombination. Clipping is a handy way to collect important slides you want to go back to later. The loxP mutants are created to avoid these problems.[23]. This recombinase comes from the P1 bacteriophage. As alluded to above, the Cre recombinase catalyzes the site specific recombination event between two loxP sites, which can be located either on the same or on separate pieces of DNA. These sites, known as loxP sequences, contain specific binding sites for Cre that surround a directional core sequence where recombination can occur. Depending on the 3. Cre-Lox recombination involves the targeting of a specific sequence of DNA and splicing it with the help of an enzyme called Cre recombinase. Tap here to review the details. The site is secure. Cre recombinase consists of 343 amino acids, which can specifically recognize Lox sites. 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. The P1 bacteriophage uses these components as part of its natural viral life-cycle, and researchers have adapted the components for use in genome manipulation. 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 . FEBS Lett. 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). 2. Thus, the orientation, and location of the loxP sites are important determinants of Read Addgene's Blog on Cre alternatives from Gerald Rubin's lab. Mol Urol. Cre-lox ? Methods for producing a maize plant by crossing hybrid maize variety X03R618CYFR with another maize plant are disclosed. Site-specific recombination (SSR) involves specific sites for the catalyzing action of special enzymes called recombinases. You will earn 10 point for each question correctly answered (140 points for 14 . [10] Tsien and his colleagues demonstrated Cre-mediated recombination can occur in the post-mitotic pyramidal neurons in the adult mouse forebrain.[11]. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. The virus, called a prophage in such a state, accomplishes this via integration and excision. Scientists have developed ways to tightly control Cre expression and to optimize Cre expression once it is induced. The Cre recombinase is an enzyme that is required for Thank you for your participation! An additional level of control can be achieved by using his Cre recombinase, engineered to reversibly activate in the presence of the estrogen analogue 4-hydroxy tamoxifen. This technique is called recombinase-mediated cassette exchange and is a very convenient and time-saving way for genetic manipulation. When such a reporter strain is mated to a Cre-expressing strain, it produces offspring in which the visible marker is expressed only in tissues with Cre activity (Figure 8). By accepting, you agree to the updated privacy policy. NIH blueprint for neuroscience: Cre driver network. What is virus associated DNA, and why do I have to order it? 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. The creloxP-mediated recombination system (the "creloxP system") is an integral experimental tool for mammalian genetics and cell biology. 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]. 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. Receive the latest news, hot plasmids, discounts and more. Cre recombinase can recognize and bind to the loxP site, Unable to load your collection due to an error, Unable to load your delegates due to an error. Studies have found that Cre recombinase is the best The temperature is 37 oC and Flp recombinase is 30. Methods for producing a maize plant by crossing hybrid maize variety X00R808 with another maize plant are disclosed. Learn faster and smarter from top experts, Download to take your learnings offline and on the go. 2023 Jan 9:2023.01.09.523268. doi: 10.1101/2023.01.09.523268. CIBN-CreC and mCherry coexpression; Light inducible; Use with plasmid 26888. By whitelisting SlideShare on your ad-blocker, you are supporting our community of content creators. Cre recombinase is a tyrosine recombinase enzyme derived from the P1 bacteriophage.The enzyme uses a topoisomerase I-like mechanism to carry out site specific recombination events. Enjoy access to millions of ebooks, audiobooks, magazines, and more from Scribd. [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]. 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. Now customize the name of a clipboard to store your clips. As Cre/ loxP . Embodiments include the seeds of soybean 4826008, the plants of soybean 4826008, to plant parts of soybean 4826008, and methods for producing a soybean plant produced by crossing soybean 4826008 with itself or with another soybean variety. These plasmids can be useful for lineage tracing, evaluating infection/transfection efficiency, selecting desired clones, and making transgenic mice. 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). This need led to the development of Cre reporter strains. [7] Combining these advances in collaboration, the laboratories of Drs. Another report using two lox variants pair but through regulating the length of DNA in one pair results in stochastic gene activation with regulated level of sparseness. Bacteriophage P1 site-specific recombination. The Cre enzyme targets loxP sites at the ends of this molecule and cyclises the genome. Models thus obtained are more likely to mimic the physiological situation. As you can imagine, breeding such mice can get a wee bit complicated. Empty backbones to regulate your gene of interest: In this subtype of loxP plasmids, reporter genes indicate which cells have undergone Cre recombination. 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. However, this approach does not provide capabilities beyond insertion, such as gene deletions or modulated gene expression. The FLP-FRT technology can be an effective alternative to Cre-lox, and has also been used in conjunction with it, allowing for two separate recombination events to be controlled in parallel. eCollection 2022. 4. Fields, Pathways 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. 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. PubMed, Gu, H., Zou, Y.R., and Rajewsky, K. 1993. * Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project Tetrad analysis, positive and negative interference, mapping through somatic Chromosome walking jumping transposon tagging map based cloning, Conversion of fibroblasts to retinal cells by transcription (final). Based on these Cre-lox recombination principles, scientists have developed constructs to activate/inactivate genes when Cre is present. Enjoy access to millions of ebooks, audiobooks, magazines, and more from Scribd. Below, I would to focus on two additional issues: using Cre/lox mice to turn on or off transgenes, and using Cre reporter strains. Two plasmids can be joined using the variant lox sites 71 and 66. 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 . This reaction synthesizes the Holliday junction discussed earlier. In our study, we mutagenized 13 bp ( 5- ATAACTTCGTATA-3) of the right RBE of the loxP sequence. the loxP sites will delete the gene 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. Learn about the latest plasmid technologies and research tools. In tissues with no cre gene the target gene with be present and function normally. In this example, expression of Cre recombinase removes the transgene (Figure 6). Weve updated our privacy policy so that we are compliant with changing global privacy regulations and to provide you with insight into the limited ways in which we use your data. We archive and distribute high quality plasmids from your colleagues. Systems, Research 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 (). sites contain nucleotide variations in the left or right arms, respectively named LE and RE. Use of the system has greatly expanded our ability to precisely interrogate gene function in the mouse, providing both spatial and temporal control of gene expression. Looks like youve clipped this slide to already. Department of Biotechnology An important application of the Cre-lox system is excision of selectable markers in gene replacement. RuvC is just one of these Resolvases that have been isolated in bacteria and yeast. [19], Cre recombinase can be synthesized by the corresponding gene under the direction of cell-specific promoters, including promoters under the control of doxycycline. Flanking a DNA sequence with loxP sites allows for the target sequence to be manipulated by Cre using a cyclization mechanism. 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. 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. Cre-lox, The cre-loxP-mediated recombination system (the "cre-loxP system") is an integral experimental tool for mammalian genetics and cell biology. Do not sell or share my personal information, 1. 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]. aegypti. I will also explain how to use Cre reporters). 1992. At a DNA level many types of manipulations can be carried out. The versatile properties of Cre recombinase make it ideal for use in many genetic engineering strategies. b) The second step is supply of Cre recombinase. Chief among these is Cre recombinase, discovered in the P1 bacteriophage and named as such because it c auses re combination (Cre) between specific DNA sequences referred to as loxP (locus of crossover (x) in P1) sites, each of which are 34 base pairs in length 1. Wang DX, Qing SL, Miao ZW, Luo HY, Tian JS, Zhang XP, Wang SN, Zhang TG, Miao CY. The Cre/lox system is one of the most powerful and versatile tools developed for mouse genetics. Transgenic mice containing a gene surrounded by loxP sites are mated with transgenic mice that have the cre gene expressing only in one cell type. Heteromeric . 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. A novel maize variety designated X00R808 and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. Application of Cre-loxP system to the urinary tract and cancer gene therapy. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). (<U>G</U>ene-<U>A</U>uto-<U>E</U>xcision <U>B</U>inary <U>S</U>ystemGAEBS)(transcription system) . A conditional knockout (cKO) model is generated via several steps, using Cre-LoxP system as an example: The targeted fragment to be knocked out is flanked by LoxP elements. 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). Phages like the lambda phage use their site specific recombinases to integrate their DNA into the host genome during lysogeny. By undergoing Cre recombination, the region floxed by the loxP sites will become inverted, [12] this process is not permanent and can be reversed. Here is the most efficient breeding scheme for generating Cre/loxtissue-specific or inducible knockouts. Plasmid Elements, doi: 10.1002/jbm4.10706. Notice that only one generation of breeding is required (Hooray!). We've encountered a problem, please try again. The protein will stay in this location in its inactivated state until tamoxifen is given. [8] They observed 50% of the DNA polymerase beta gene was deleted in T cells based on DNA blotting. How can I be notified when a plasmid from a specific lab or paper is available? The Cre or cre-recombinase protein (called so because it causes recombination) catalyzes site-specific recombination events between two DNA recognition sites. 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. The points where the integration and excision reactions occur are called the attachment (att) sites. 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. Furthermore, recombination systems such as Cre-lox are widely used, mostly in eukaryotes. [13] Mechanism of translocation [ edit] More novel and sophisticated uses for the Cre/loxsystem are being developed all the time. In this context we created four target site lines for RMCE and evaluated their fitness costs. 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. It allows the DNA modification to be targeted to a specific cell type or be triggered by a specific external stimulus. 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. IHDR y ] + sRGB pHYs od FIDATx^.{s" ""S If you run into any problems registering, depositing, or ordering please contact us at [emailprotected] The obvious difference between the two systems is that the recombinase (Cre and Flp) has different optimal reaction temperatures. of interest. A soybean cultivar designated 03070412 is disclosed. In fact, the enzyme-DNA complex is an intermediate stage, which is followed by the ligation of the 3 OH group of one DNA strand to the 5 phosphate group of the other DNA strand, which is covalently bonded to the tyrosine residue; that is, the covalent linkage between 5 end and tyrosine residue is broken. 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. Commonly used gene replacement strategies introduce selectable markers into the genome to facilitate selection of genetic mutations that may cause growth retardation. Initiation of site-specific recombination begins with the binding of recombination proteins to their respective DNA targets. Missirlis PI, Smailus DE, Holt RA. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. This provides the advantage that the Cre recombinase is active for a short time. 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 . 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. If appropriate for your experiments, you can maintain a colony that is homozygous for theloxPflanked allele, but in which one parent was hemizygous/heterozygous forcreand the other was a noncarrier/wild-type forcre. place. The limitations of creloxP technology and related considerations for experimental design are addressed, and alternative strategies for sitespecific genetic recombination and integration are discussed. Cre recombinase removes the stop sequence, so the transgene is expressed only in the cells where Cre is active (see Figure 4 below). First, the nucleotide sequence identity in the spacer region of lox site. FOIA 2022 Dec 21;7(1):e10706. The result of recombination depends on the orientation of the loxP sites. 5 Phosphate ends are usually the substrates for this reaction, thus extended 3 regions remain. GENE KNOCKOUT USING THE CRE-LOXP SYSTEM For a few examples of such strains, check out 008179 and 006481. Nov 2, 2015. Once tamoxifen is introduced, it is metabolized into 4-hydroxytamoxifen, which then binds to the ER and results in the translocation of the CreER into the nucleus, where it is then able to cleave the lox sites. Cre (Cre recombinase) Cre is a 38 kDa recombinase protein from bacteriophage P1 that catalyzes recombination between LoxP sites. By PMC It can transition to the lytic cycle by a later event such as UV radiation or starvation. Addgene's viral service provides high-quality and high-titer viral preparations of plasmids that are currently in our repository. The exact sequence is given below; 'N' indicates bases which may vary, and lowercase letters indicate bases that have been mutated from the wild-type. The P1 plasmid is relatively large (90Kbp) and hence exists in a low copy number - usually one per cell. Plasmids. loxP (locus of X-over P1) is a site on the bacteriophage P1 consisting of 34 bp. The SlideShare family just got bigger. The Cre recombinase can recognize cryptic sites in the host genome and induce unauthorized recombination, damaging host DNA. At the given specific site on the DNA, the hydroxyl group of the tyrosine in the recombinase attacks a phosphate group in the DNA backbone using a direct transesterification mechanism. Maize hybrid X13C737 is an invention by Andrew Jon Ross, Lisbon IA UNITED STATES. 3. The Cre-loxP recombination system is routinely used for the generation of mouse knockouts. The Cre/lox system can be used to turn off a transgene. This has been largely due to . conditional genetics; cre; cre-loxP; gene editing; loxP; recombination; site directed mutagenesis. Simply put, the Cre/loxsystem rocks. Can be used for injection into fertilized zygotes, Cre activates gene of interest by removing neoR and stop sequence; used for making transgenic mice. The Cre protein (encoded by the locus originally named as "Causes recombination", with "Cyclization recombinase" being found in some references)[21][22] consists of 4 subunits and two domains: The larger carboxyl (C-terminal) domain, and smaller amino (N-terminal) domain. [9] Incomplete deletion by Cre recombinase is not uncommon in cells when two copies of floxed sequences exist, and allows the formation and study of chimeric tissues. mouse), and another with the loxP sites (the floxed mouse) are created, and crossed to. A novel maize variety designated X03R618CYFR and seed, plants and plant parts thereof are produced by crossing inbred maize varieties. 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). These targeted DNA changes are useful in cell lineage tracing and when mutants are lethal if expressed globally. The use of the Cre/loxP system for generating tissue specific or inducible knockouts is a powerful tool for mouse genetics, but it is only one piece of the total picture. This patent application was filed with the USPTO on Wednesday, February 15, 2012 It was unclear whether only one allele in each T-cell or 50% of T cells had 100% deletion in both alleles. Cre/lox switchingexpression system Currently, mostwidely used site-specific DNA recombination system system[18, 19]. bioRxiv. Applied Biological Materials - abm 40.7K subscribers Subscribe 1.4K 102K views 3 years ago The FLEx vector or "flip. Matsumura T, Watanabe T, Totsugawa T, et al. FEBS Lett. 18 In some cases a genomic variant can be present but not detected by the, With November designated as Family Health History month, clinicians are sure to have seen lots of messaging, The microbiome plays a vital role in our health and wellbeing. As with any other technology or research tool, it has limitations and pitfalls that need to be considered while planning experiments or interpreting results. A number of conserved site-specific recombination systems have been described in both prokaryotic and eukaryotic organisms. The Cre protein is a site-specific DNA recombinase that can catalyse the recombination of DNA between specific sites in a DNA molecule. Many Cre constructs also contain fluorescent labels that indicate if recombination has occurred, allowing for direct comparison of Cre+ and Cre- cells. 1992. Two factors have been shown to affect the efficiency of Cre's excision on the lox pair. The activity of the Cre enzyme can be controlled so that it is expressed in a particular cell type or triggered by an external stimulus like a chemical signal or a heat shock. 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]. Again, only one generation of breeding is required. government site. The circular molecules contain site-specific recombination sequences and allow transformation of host cells with DNA comprising only selected sequences of interest. [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. 8600 Rockville Pike 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). Before Both AAV and Lenti preps of Cre are available. This can also take place in the absence of the Cre lox system[39] with the help of other bacterial and viral proteins. Therefore, researchers often use transgenic mice expressing CreERt2 recombinase induced by tamoxifen administration, under the control of a promoter of a gene that marks the specific cell type of interest, with a Cre-dependent fluorescent protein reporter. The Cre-lox system is used to introduce deletions, insertions, translocations, and inversions at a specific genomic locus. 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. This breeding scheme may not be the most efficient one for producing all Cre/loxstrains. Assistant Professor (Ad Hoc) Have you used the Cre-lox system in your research? the Flp-FRT system. Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. Such conditional lineage tracing has proved to be extremely useful to efficiently and specifically identify vascular smooth muscle cells (VSMCs) and VSMC-derived cells and has been used to test effects on VSMC and VSMC-derived cells in vivo.[32][33][34][35][36][37]. Cre-Lox recombination is a site-specific recombinase technology widely used to carry out deletions, insertions, translocations and inversions in the DNA of cells. In part 1 of this mini-series, I introduced the concept and applications of Cre-loxP. This website uses cookies to ensure you get the best experience. It appears that you have an ad-blocker running. . Unauthorized use of these marks is strictly prohibited. Kim H, Kim M, Im SK, Fang S. Mouse Cre-LoxP system: general principles to determine tissue-specific roles of target genes. A novel maize variety designated 2ZZSB1121 and seed, plants and plant parts thereof are provided. These constructs allow for Cre-regulated gene expression. (Already know this? Since homologous recombination occurs after DNA replication, two strands of DNA are available, and thus, the 3 OH group must pair with its complement, and it does so, with an intact strand on the other duplex. It appears that you have an ad-blocker running. 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.
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