...help of Scott Lindner . Cut Fully functional CRISPR/Cas enzymes will introduce a double-strand break (DSB...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...research for decades. Recent developments with CRISPR-Cas9 gene editing have propelled both reverse and forward...forward genetics of these parasites. Moreover, CRISPR-dCas9 gene regulation has also been established for some...

...more information on the wide variety of Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems...species or PAM binding variant of Cas9. For instance, wild-type SpCas9 must be used with targets that are...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...increases off-target effect. Which species or variant of Cas9 ( S. pyogenes , S. aureus etc.) was this gRNA sequence...

...Lab General Cloning Protocol 237.2 KB SpCas9 Plasmids The CRISPR/Cas system can be implemented in mammalian...Return to top References SpCas9 plasmids Multiplex Genome Engineering using CRISPR/Cas Systems. Cong L, Ran...hosts contain a combination of CRISPR-associated (Cas) genes as well as non-coding RNA elements capable...efficiency. SpCas9 and SpCas9n with 2a-Puro and 2a-EGFP are also available. 2. SpCas9 (or SpCas9n, D10A nickase...Protocols 2013). SpCas9 Cas9: The PX551 plasmid (AAV-SpCas9, #60957) contains a single cassette expressing ...plasmid expressing SpCas9 alone without sgRNA. Full references are below. 1. SpCas9 (or SpCas9n, D10A nickase...plasmids contain two expression cassettes, a human codon-optimized SpCas9 or SpCas9n, and the single guide RNA...

...Sensor for Ex Vivo and In Vivo Imaging of Ca Dynamics in High-Ca Organelles. Cell Chem Biol. 2016 Jun 1....Calcium erGAP3 (GFP-Aequorin Protein) for imaging of Ca+ dynamics in endoplasmic reticulum GFP-Aequorin Protein...calcium sensor T-GECI Blue-shifted genetically encoded Ca(2+) indicator with enhanced two-photon absorption...green, red or blue/green) Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA. Nat Commun...fluorescent CEPIA indicators for visualization of Ca(2+) dynamics in mitochondria. Sci Rep. 2020 Feb 18...CEPIA1er ER-mitochondria tethering by PDZD8 regulates Ca(2+) dynamics in mammalian neurons. Science. 2017 ...Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins...

...puromycin resistance cassette, such as PX459 ). After you co-transfect the CRISPR/Cas system with your homologous...that can be inserted into the genome using CRISPR/Cas for Homologous Recombination (HR)? How long should...the Cas9 nickase doesn't have the ability to induce double-stranded breaks like the wildtype Cas9 does... engineering experiments, consider that wildtype Cas9 with optimized chimeric gRNA has high efficiency...effect). The double nickase system is based on the Cas9 D10A nickase described in Figure 4 of the Cong, ...express two spacers and use PX335 to express the Cas9n (nickase). The concept of the double nickase system...can express two different chimeric gRNAs with the Cas9 nickase which will together introduce cleavage of...

...found at the Goldstein Lab website . SapTrap CRISPR/Cas Toolkit - Erik Jorgensen Lab. A collection of plasmids... via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses purified Cas9, such as that...feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific transgenic...worm CRISPR resources include plasmids expressing Cas9, empty gRNA backbones, and constructs for endogenous...that incorporate a self-excising drug selection cassette (SEC). More information regarding this kit can...targeting vectors for tag insertion using CRISPR/Cas9 and the selection of C. elegans strains. CRISPR ...that expressed from pHO4d-Cas9 from Michael Nonet. Protocol can be found at the (Link opens in a new window...

...Targeted Integration Using CRISPR/Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression...Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9 Vector System...modified CRISPR-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference...morpholinos, zinc finger nucleases, TALENS, and CRISPR/Cas9-based gene editing make the zebrafish a stalwart...resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of useful...Knock-in tagging - Michel Bagnat lab. Targeting cassettes encoding fluorescent proteins for tagging genes...use of zebrafish. CRISPRz - A database of CRISPR/Cas9 target sequences that have been experimentally validated...

...Ciliary Beating Affected by the Pcp4 Dose-Dependent [Ca 2+] i Increase in Down Syndrome Mice, Ts1Rhr . Do...infection. General Tools : Plasmids expressing Cas12a/Cas13a, Reverse Transcriptases, etc. that may be useful...new window) Find Zhang Lab Cas13a plasmids Additional protocol for using Cas13a ( pC013 ) for detection ... has utilized the natural RNAse activity of the Cas13 protein to develop and optimize the method termed...coronavirus - Enabling coronavirus detection using CRISPR-Cas13: Open-access SHERLOCK research protocols and design...SARS-CoV-2: A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic Material. ...2020.04.20.052159 . Jennifer Doudna's lab has utilized Cas12a’s non-specific ssDNA degradation to develop the ...

... PubMed (Link opens in a new window) Marcinkiewcz CA, Mazzone CM, D'Agostino G, Halladay LR, Hardaway ...

...Candido TR, Castellarin M, Chen J, Chen Y, Cheng JC, Chopra V, Docking TR, Dreolini L, D'Souza CA, Flynn EK...webpages. These plasmids are described in Portales-Casamar et al., 2010 . List of Pleiades MiniPromoters MiniPromoter...intron-lacZ/NLS Additional Resources References Portales-Casamar E, Swanson DJ, Liu L, de Leeuw CN, Banks KG, Ho...