...Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4...also be used to design primers to read states by qPCR or Sanger sequencing as described in the Material...hammerhead ribozyme parts are described in Nielsen et al (3) and Lou et al (4). Despite choosing proper register...machine design space is highly degenerate and often times there are multiple register designs that may...recognition sites from the register. If the RSM is designed in such a way that each possible order of inputs...segments in each state of a register enables the design of primer pairs that PCR amplify in some states...registers within a sample using a metric that we describe in the Materials and Methods section of our paper...

...bacterial mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector...on channelrhodopsins – Chronos & Chrimson Through de novo sequencing of 127 algal transcriptomes, as well...components into a destination vector in a predefined order. Finally, the destination vectors are used ...engineering approach, they designed Opto-RTKs, which activate signaling cascades on exposure to low-intensity...years of distributing his Designer Receptors Exclusively Activated by Designer Drugs (DREADD) plasmids ...minimizing confounding secondary effects? An approach described as “knocksideways” (a British idiom for “taking...recombination elements such as FRT or Lox sites, and provides a modular system for easier exchange and re-use...

...applications. LuxSit-i : An artificial luciferase desined de novo using deep-learning approaches. Split-Luciferase...to that of the fluorophore. Conformationally destabilized Firefly luciferase mutants : A set of three ...three sensors that are increasingly structurally destabilized versions of Firefly luciferase which serves ...dimerization , endosomal escape of cell-penetrating peptides , and more. nLuc/cLuc for split Firefly Luciferase...mammalian systems. ID Plasmid Luciferase Type(s) Description PI 16539 pBV-Luc Firefly Insertion of 5' promoter...service icon . ID Plasmid Luciferase Type Promoter Description PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc...your regulatory element of interest. ID Plasmid Description Gene/Insert PI Return to Top Do you have suggestions...

...X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. 2014. Genome-Scale CRISPR-Cas9...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR revolution shows... store a record of invading phages but also to destroy the phages upon re-exposure. More specifically,...complex binds to the foreign DNA and cleaves it to destroy the invader ( Figure 1 ). Figure 1: An overview...in vitro. In 2013, Cong et al. and Mali et al. described CRISPR-based genome editing in mammalian cell ... of new applications. The first CRISPR papers described two main categories of genome edits. First, the...very low off-target activity due to rationally designed mutations. Researchers have expanded the CRISPR...

..., Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Science . 2014 Jan 3;343(6166...oligos (design is indicated below) can be cloned into the CRISPR array. The oligos are designed based on...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Jump to: SpCas9 Plasmids...digested using BbsI, and a pair of annealed oligos (design is indicated below) can be cloned scarlessly into...vector before the sgRNA scaffold. The oligos are designed based on the target site sequence (20bp) and need...using SAM: Addgene plasmids #61422 - #61424 were designed for transient transfection : 61422 : Expresses... references are below. Available plasmids are described below: 61591 : PX601; CMV-driven SaCas9; U6-driven...

...Strausbaugh LD, Gray P, Vartak NB, Brown M, Talbot D, Berg DE. pBR322-derived multicopy plasmids harboring large...confirmations like linear and nicked plasmids, as described in this article.   Long-read sequencing Addgene...bacterial colonies using one of the detection methods described above can help ensure the isolation of the monomeric...copy number plasmids causes instability: CoIE1 encodes a determinant essential for plasmid monomerization...Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design: impact on efficacy, safety and upstream production...

... cell reprogramming factors and wait for cells to de-differentiate. However it may be difficult to decide...blog post Delivery Methods for Generating iPSCs provides an overview of several different reprogramming...find more information. Delivery Method Species Description Article PI Lentivirus Human Expression of human...

...https://doi.org/10.1093/nar/gku289 Chen C, Fenk LA, de Bono M (2013) Efficient genome editing in Caenorhabditis... DNA double-strand breaks up to 54 bp from the desired insertion site result in efficient editing. Repair...available through Addgene. In 2019, Farboud et al. described another similar co-conversion marker (repair of...whether it be by improved gRNA or repair template design, delivery, screening, and more. Here are some highlights...highlights from the past few years:  Repair template design The Meyer lab meticulously characterized several...of editing. First, they devised guidelines for designing single-stranded repair templates (Farboud et al...Tools such as SapTrap Builder greatly simplifies design of these constructs, allowing novice researchers...

...policies that “muzzle” government scientists to the de-funding of crucial environmental research programs...and minority communities, where these issues hit hardest. According to the Guttmacher Institute, most women...was supremely proud of Nashville that morning. Despite the rain, about 4,000 supporters of science showed... an agenda that halts current data collection, destroys existing data, and guts future funding.I marched...science belongs to the taxpayers who fund it, and we deserve to know the truth about our world. Jonathan Aguilar...

... and B.Z. Stanger, Adult cell plasticity in vivo: de-differentiation and transdifferentiation are back...

... cell reprogramming factors and wait for cells to de-differentiate. However it’s difficult, especially... with peptides that help mediate their transduction, such as the polyarginine peptide described in Zhou...reprogrammer, to decide which method to use. This post provides a brief overview of reprogramming methods with... Paramyxoviridae family of viruses, which also includes measles and mumps. Sendai transduces a wide range...in practice integration can occur. Okita et al describes a protocol for generating iPSCs by plasmid transfection...

...Off-Spotter: tool for CRISPR/Cas design Program for designing optimal gRNAs. Provides feedback on number of potential...the Chan Zuckerberg Biohub . gRNA Design Deep learning models to design gRNAs and predict their efficiency... a new window) : a webtool that provides models for SpCas9 gRNA design. DeepBE (Link opens in a new window...window) : a webtool that provides models for base editor (BE) gRNA design. DeepPrime (Link opens in a ...window) : a webtool that provides models for prime editor (PE) pegRNA design. PMID 37119812 (Link opens...purpose of designing and evaluating experiments. Green Listed - a CRISPR Screen Tool Design gRNAs for ...Eukaryotic Pathogen CRISPR guide RNA/DNA Design Tool (EuPaGDT) gRNA design tool with extensive selection of eukaryotic...

...Sangamo)-Destination Pawel Pelczar pCAG-T7-TALEN(Sangamo)-Destination constructs were designed for optimal...pC-GoldyTALEN and RCIsricpt-GoldyTALEN are designed as destination vectors for the Voytas lab Golden Gate ...Plasmids pTAL5-BB and pTAL6-BB are designed as alternative destination vectors for the Golden Gate TALEN...TALEN(Sangamo)-FokI-KKR-Destination 40132 pCAG-T7-TALEN(Sangamo)-FokI-ELD-Destination Golden Gate TALEN 2.0...Mendenhall The TALE plasmid vectors described on this page are designed to target and edit the epigenome ...with custom repeat arrays. TALEN Kits TALEN Kit Description Depositing PI FusX TALEN Assembly System Contains... pre-assembled trimer and tetramer RVD domains designed for rapid TALEN assembly. Kiran Musunuru and Chad...

...more about how to design your gRNA . Browse Plasmids: gRNAs Synthesize and clone desired gRNAs Once you ...sequences it is time to design your gRNA oligos and clone these oligos into your desired vector. In many cases... Science Guides CRISPR Guide Plan Your Experiment Plan Your CRISPR Experiment Jump To: Plan Your Experiment...Select Genetic Manipulation Select Expression System Design gRNA Publications Get Started CRISPR is a powerful...available for your particular experiment. Select Your Desired Genetic Manipulation Different genetic manipulations...mammalian cell lines Select Your Target Sequence and Design Your gRNA Once you have selected your CRISPR components..., you are ready to select a target sequence and design your gRNA. Know your cell line/organism and genomic...

...more. CRISPR Design FAQs Homologous Recombination (HR) FAQs At the Bench FAQs CRISPR Design FAQs Should...template vector design guidelines? A few notes below are considerations for designing HR donor. Generally...usually pick 3-6 guides around the region to find the most efficient guide (most guides would work, but... this ‘split RNA’ design is not as efficient in genome editing as the second design (in pX330 / PX335 ...comparable efficiency to the optimized chimeric design but with better accuracy (in other words, lower...nickase system is based on the Cas9 D10A nickase described in Figure 4 of the Cong, et. al, 2013 Science ...published with thorough step-wise instructions on designing and using Cas9-based genome engineering tools ...

...Membrane Localization pWZL-Neo-Myr-Flag-DEST - pDest vector that adds a N-terminal myristoylation...expression pLenti CMV Neo DEST - Lentiviral Gateway destination vector for gene ...expression pLenti CMV Puro DEST - Lentiviral Gateway destination vector for gene ...expression pLenti CMV Hygro DEST - Lentiviral Gateway destination vector for gene ...Mammalian pLenti X1 GFP-Zeo DEST - Lentiviral Gateway destination for shRNA ...pLenti CMV/TO Zeo DEST - Tet-inducible lentiviral Gateway destination vector ... delivery pLenti CMV GFP DEST - Lentiviral Gateway destination vector for gene ...

... pii: 56469. Edwin Chapman Neuropeptides GRAB biosensors for neuropeptides VIP, NPY, CCK, NTS, UCN, SST... orexin neuropeptides A genetically encoded sensor for in vivo imaging of orexin neuropeptides. Nat Methods...Biosensors Fluorescent biosensors are proteins designed so that their fluorescent properties are sensitive... or organisms for long-term imaging and can be designed to specifically localize to cellular compartments...compartments or organelles. Addgene’s database includes many fluorescent biosensors that signal the presence ...environmental conditions. With proper experimental design, biosensors can enable investigation of a signaling...with the viral service icon . Metal Ions Target Description Article PI Calcium jGCaMP8 Fast Genetically Encoded...

...contains many plasmid tools that were specifically designed for use in plants. These resources include tools... to fluorescent proteins. Addgene’s collection includes plasmids containing promoters that drive general... Chen. Cas9 and gRNA backbone for use in rice. Includes a protocol (Link opens in a new window) for Targeted... Yang. The MoClo CRISPR/Cas Toolkit for Plants includes CRISPR/Cas nucleases, base editors, gRNA backbones...Nekrasov. The MoClo-compatible zCas9i Cloning Kit includes a high efficiency intron-optimized SpCas9-coding...Caixia Gao. Cloning The following collections are designed for creating plant expression vectors: A modular...MoChlo: Modular Cloning Chloroplast Toolbox for design of synthetic operons for chloroplast transformation...

...Table 4. Gateway Destination Vector ID Plasmid 48222 pAC90-pmax-DEST : Gateway Destination vector derived...the dCas9 protein provides the transactivation activity, the sgRNA component provides the specificity and...and can be transferred to Gateway destination vectors (e.g., pmax-DEST [48222] and (Link opens in a new...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts CRISPR-on is an RNA-guided... and is designed to target promoters or enhancers of genes of interest. Different flavors of activators...new window) Lifetechnologies Gateway Destination Vectors ) to adapt to different systems by LR clonase ...clonase reaction. These plasmids are described in: Multiplexed activation of endogenous genes by CRISPR-on, ...

...the biology of C. elegans and other nematodes. Wormbuilder - Describes methods to engineer the genome of...characterized which allow researchers to express desired constructs in precise cell types and/or developmental...that contain tools for use in worm research. This includes CRISPR, RNAi, empty backbones, fluorescent proteins...expression plasmids. Large collections or kits are described below and not included in this table. For a more...search Addgene's entire collection. ID Plasmid Description Gene/Insert PI Plasmids Expressing Worm Genes...strains. CRISPR plasmids from John Calarco's lab as described in Heritable genome editing in C. elegans via ...genome-editing events. Developed in Craig Mello's lab and described in A Co-CRISPR Strategy for Efficient Genome Editing...