...References Portales-Casamar E, Swanson DJ, Liu L, de Leeuw CN, Banks KG, Ho Sui SJ, Fulton DL, Ali J, ...Plasmid Collections Pleiades Promoter Plasmids Pleiades Promoter Project The Pleiades Promoter Project (... These plasmids are described in Portales-Casamar et al., 2010 . List of Pleiades MiniPromoters MiniPromoter...Plasmids from the Pleiades Promoter Project - A regulatory toolbox of MiniPromoters to drive selective... through these human MiniPromoters. Browse the Pleiades Promoter Plasmids using the table below. Additional...

...activities. By using GFP as a scaffold, scientists can design de novo systems using modular components or take ...

...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense), and four transposase components (TnsA, TnsB...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts CRISPR-associated transposases...

...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex, comprised of a combination of Cas5...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Cas3 is a unique member...

...Deletions in Mammalian Cell Lines via CRISPR/Cas9. Bauer DE, Canver MC, Orkin SH. J. Vis. Exp. (95), e52118, ...Cas9 in mammalian cells. Canver MC, Bauer DE, Dass A, Yien YY, Chung J, Masuda T, Maeda T, Paw...manuscript PDF posted at the top of the page. CRISPR Design Design sgRNAs manually or using freely available online... be 24-mer oligos ( Table 4 ). Design Deletion Screening Primers Design one set of primers internal to...Here we present a simple methodology for CRISPR design, cloning, and delivery for the production of genomic...genomic deletions. In addition, we describe techniques for deletion, identification, and characterization...

...(Link opens in a new window) References Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B. In vivo...

... Hacohen N, Hahn WC, Lander ES, Sabatini DM, Root DE. Cell. 2006. 124:1283-1298. PubMed . A versatile ...site. Find resources for more information on shRNA design and delivery. Constitutive Lentiviral shRNA Lentiviral...for constitutive shRNA expression ID Plasmid Description PI 10878 pLKO.1 ‐ TRC cloning vector shRNA expression... included in the vector to monitor expression; Designed for inducing RNAi in a wide range of cell types...conditional (Cre-lox) shRNA expression ID Plasmid Description PI 11578 pSico Cre addition causes EGFP to be...for Tet-regulated shRNA expression ID Plasmid Description Tet on or off PI 11643 pLVCT-tTR-KRAB 2nd generation...for constitutive shRNA expression ID Plasmid Description PI 8452 pMKO.1 puro Retroviral vector for shRNA...

...Stedingk K, Fujita T, Persson CU, Eriksson P, Veerla S, De Preter K, Speleman F, Fujii H, Påhlman S, Mohlin ...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Originally developed in...can be ordered via the links below: ID Plasmid Description 47948 3xFLAG-dCas9/pCMV-7.1 enChIP plasmid, expresses...

...M. M., Semenova, E., Severinov, K., De Vos, W. M., Dame, R. T., De Vries, R., Brouns, S. J., & Van Der...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex to initiate DNA degradation. Cascade...Cascade C RISPR- as sociated c omplex for a ntiviral de fense; a complex of multiple Cas enzymes including...Han, Y., Ogrodowski, P., Crippa, A., Rekhtman, N., De Stanchina, E., Lowe, S. W., & Ventura, A. (2014)...., Boix, C., Perrouault, L., Tesson, L., Geny, S., De Cian, A., Itier, J. M., Anegon, I., Lopez, B., Giovannangeli... the desired HDR-edited allele. Therefore, it is important to confirm the presence of the desired edit..., the longer pegRNA also encodes a primer binding site (PBS) and the desired edits on a RT template. The...

...none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm EcoRI none S. pyogenes de Bono pMB60 47941...pyogenes LEU2 Lu pDestTol2CG2-U6:gRNA 63156 Zebrafish BseRI none S. pyogenes Zon pDestTol2pA2-U6:gRNA 63157...module vectors, including three designed for dicots and three designed for monocots. Using these gRNA ...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts CRISPR requires that you...Cas9 (or other CRISPR) system that the vector was designed to be used with, such as S. pyogenes, S. aureus...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...from our collection. Plasmids Expression System Description PI Multiplex CRISPR/Cas9 Assembly System Kit ...

...in transcriptional repression, activity dependent de-repression, chromatin remodeling, gene activation...species (mouse or human) Mutation ID Plasmid Description Gene/Insert Mutations PI Return to top External...BrainSpan Atlas of the Developing Human Brain - includes developmental transcriptome information, prenatal...

...Pietro De Camilli 194005 VPS13D1-1576-(Y1253S) VPS13D GFP CMV Spinocerebellar Ataxia Pietro De Camilli...145 SYNJ1 Flag CMV Parkinsonism, early-onset Pietro De Camilli 22292 pcDNA3-FLAG-Synaptojanin 1-170 SYNJ1...SYNJ1 Flag CMV Parkinsonism, early-onset Pietro De Camilli 22293 pEGFPC1-FLAG-Synaptojanin 1-145 SYNJ1 GFP... GFP, Flag CMV Parkinsonism, early-onset Pietro De Camilli 22294 pEGFPC1-FLAG-Synaptojanin 1-170 SYNJ1...SYNJ1 GFP, Flag CMV Parkinsonism, early-onset Pietro De Camilli 22563 Flag-HA-UCHL1 UCHL1 HA, FLAG Parkinson's...inserted at position aa 1914 CMV Parkinsonism Pietro De Camilli 118791 pHBS1383 TDP43 RRM-GFP FYW-S TARDBP...delta1235-1748 VPS13C Halo CMV Parkinsonism Pietro De Camilli 187767 MAC_C_CSF1R CSF1R MAC Hereditary diffuse...

...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...

... 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...

...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 ...