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We narrowed to 53 results for: stem loop

Showing: 21 - 40 of 53 results

  1. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ... the ENCODE project, find enhancers in embryonic stem cells, and conclude with a targeted CRISPR screen..., an essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds just over 13,000...reveals which NANOG-related genes are critical to stem cell survival. Knowing which genes influence the...screen. We start by infecting millions of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated...step 2. The result is a heterogenous population of stem cells, usually housed in a single flask. Through...enhancer it targets is a “pillar of function” for our stem cells. Remove this pillar and the ESC dies. Order...of which stretches of the genome are pillars for stem cells, or metastasized tumor cells, or alzheimers-affected...

  2. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ... ssDNA secondary structure — for example, the stem-loop structure of transcriptional terminators. Another...cloning) would be a true rebel. Not only does this system not use site-specific recombination, it also doesn...’t require a ligation step. Based on the robust system of homologous recombination found in E. coli, SLIC...

  3. Zhang Lab CRISPR Page

    Type
    Collection
    ... : sgRNA cloning backbone with MS2 loops at tetraloop and stemloop - Contains BbsI sites for insertion...lentiviral sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker. Contains... formats: 1 vector system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9-Blast ...incorporating two MS2 RNA aptamers at the tetraloop and stemloop 2 The MS2-P65-HSF1 activation helper protein... P alindromic R epeats) is a microbial nuclease system involved in defense against invading phages and.... We have harnessed the type II CRISPR nuclease system to facilitate genome editing in mammalian cells...Protocol 237.2 KB SpCas9 Plasmids The CRISPR/Cas system can be implemented in mammalian cells by co-expressing...

  4. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ...cytokines are a potent inducer of an autoinflammatory loop: keratinocytes both respond to and release IL-36...aortic disease,” Prakash says. In induced pluripotent stem cells derived endothelial cells from women who are...

  5. Hot Plasmids - August 2020

    Type
    Blog Post
    ...new multiplexed CRISPRi and CRISPRa system for human pluripotent stem cells By Jennifer Tsang CRISPR gene...To overcome these variations in human pluripotent stem cells(hPSCs), Lindy Barrett’s lab at the Broad Institute..., stripe pattern-forming incoherent feed-forward loop. Find the plasmids at Addgene! New from the...improve utility of the system for use with low affinity variants of TEV. The new system has been demonstrated...cells Nanobody purification CRISPRi and CRISPRa system for hPSCs The CRISPR corner New from the viral ...studies.  Find the plasmids to set up your screening systems at Addgene! Schematic of the evolution platform...piggyBac plasmids include a mRFP reporter. With this system, they could quantify and track cells expressing...

  9. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...that aptamer insertion in the gRNA tetraloop (a four base hairpin loop motif within the gRNA) resulted in...School developed a dCas9-based multicolor labeling system to visualize chromosomal loci in human cells (Ma...sites on different chromosomes in living cells, the system requires high-copy chromosome-specific loci in ...stability. First, they began with the Broccoli aptamer system to visualize the gRNA in living cells. Broccoli...inserted an octet of MS2 hairpins into the gRNA tetraloop and went through subsequent design iterations ...measure inter-locus distances. They tested this system out on chromosome 19 by targeting 46 repetitive...

  10. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ...serotypes that vary in their RBE and T-shaped hairpin loop sequences (Wilmott et al. 2019). Most ITR sequences...Viral Vectors 101: Pseudotyping Viral Vectors 101: Systemic Capsids More resource on Addgene.org Addgene's...rAAV8 Production Using the Baculovirus/Sf9 Cells System. Hum Gene Ther Methods, 28(5), 277–289. https:/...

  11. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...with the Cas13a molecule through a uracil-rich stem loop and facilitates target binding and cleavage through...Abudayyeh et al., 2016). In the endogenous CRISPR/Cas9 system, Cas9 uses both a tracrRNA and crRNA to facilitate...high specificity (Gootenberg et al., 2017). This system, dubbed SHERLOCK (depicted in figure 1) has been...specific PFS, making it a very flexible cleavage system. LwaCas13a is also amenable to multiplexing; expressing...characterization of diverse class 2 CRISPR-Cas systems." Molecular cell 60.3 (2015): 385-397. PubMed PMID...

  12. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...and Doench labs utilized tracrRNAs with two PP7 stem loops to recruit additional transcription factors to...for vemurafenib resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic Activation Mediator) uses...well as more difficult conditions such as model systems and primary cells. References 1. Doench, John ...Tanenbaum, Marvin E., et al. "A protein-tagging system for signal amplification in gene expression and...

  13. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...simpler in structure than Cas9’s; only a short stem-loop structure in the direct repeat region is necessary...and tested? Class 2 CRISPR systems, including the type II Cas9-based system, require a single-component...Cas9 Type II CRISPR systems based on Cas9 were thought to be the simplest CRISPR systems and the easiest ...RNA-guided endonuclease of a class 2 CRISPR-Cas system. Figure from Zetsche et al., 2015. How was ... the multi-subunit complex employed by class 1 systems. A putative new class 2 nuclease, Cpf1 (CRISPR ...several genomes and is classified as a type V CRISPR system. Like Cas9, Cpf1 contains a RuvC-like endonuclease...this information, they turned to a cell culture system to see if any Cpf1 nucleases would exhibit in vivo...

  14. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...photo-sensitive LOV2 domain from A. sativa into a loop region of the Acr protein AcrIIA4. In the dark, ...(Acr) proteins potently inhibit type II CRISPR systems including the popular S. pyogenes Cas9 (Rauch et...light-sensitive protein domains, they have developed a system to control CRISPR-Cas9 activity using blue light...took the light-sensitive protein QuasAr2 and systematically mutated it. Using a robotic picking approach...interaction studies easier with a 2in1 vector cloning system Article contributed by Michelle Cronin  Listen ...Grefen lab introduced the “2in1” vector cloning system - a Gateway compatible approach to simultaneously...be cloned into an entry vector. The 2in1 cloning system contains two entry vectors (pUC57-L1L4 and pUC57...

  15. dTAG - You're it!

    Type
    Blog Post
    ... Is a Structural Regulator of Enhancer-Promoter Loops. Cell. December 14 2017. 171(7):1573-1588.e28 PubMed...cell-based assays and mouse models. The dTAG system The dTAG system pairs highly selective small molecule FKBP12F36V...contains the full description of the dTAG system. The dTAG system was employed to evaluate degradation of...which we refer to as the degradation tag (dTAG) system, to co-opt the endogenous ubiquitin proteasome ...protein. We are particularly excited about the dTAG system as we can use it to evaluate the effects of rapid...traditional genetic approaches. In addition, the dTAG system is versatile and can be used in both cell-based...novartis.com). Considerations when getting the dTAG system up and running in your lab Confirm the functionality...

  17. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...target various RNA molecules - such as MS2 stem loops, PPL stem loops, the Spinach2 aptamer, and more - to ...Baby Spinach core embedded in a single, extended stem-loop, contiguous with the sgRNA core (see image above...activation is capable of promoting induced-pluripotent stem cells (iPSCs) to take on a “neuron-like” phenotype...organelles, a cell cycle indicator for mouse embryonic stem cells and a series of inducible promoter constructs...CRISPR-Cas9 system to create a tool that is quick, versatile, and robust. Dubbed the LACE system for light-activated...via the CRISPR system. The ease and flexibility of the LACE technology makes this system widely accessible...gene expression by creating the SunTag system. The SunTag system, named after the "stellar explosion SUperNova...

  18. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...protein that specifically detects a specific RNA stem loop-structure (Querido and Chartrand, 2008). Finally...naturally occurring co-factor binding proteins, systems wherein a protein binds a synthetic, fluorescent...strategies works best for a specific biological system under anaerobic conditions needs to be examined...covalently label the peptide with a fluorophore. Both systems are a hybrid between a genetically encoded tag ...protein labelling in vitro and in vivo." Molecular BioSystems 12.6 (2016): 1731-1745. PubMed PMID: 26960991...in living human cells by an optimized CRISPR/Cas system." Cell 155.7 (2013): 1479-1491. PubMed PMID: 24360272...
Showing: 21 - 40 of 53 results