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We narrowed to 714 results for: CaS

Showing: 141 - 160 of 714 results

  1. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...Yang, J. et al. (7) wished to trigger intracellular Ca++ release with an optogenetic receptor, melanopsin...composed of Venus fluorescent protein, the Troponin C Ca++ binding domain, and NanoLuc® Luciferase. CalFlux...stimulation with NanoLuc-based luminescence (BRET) Ca++ sensing. Nature Comm. 7, 13268. PubMed PMID: 27786307...endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc is very inviting for endogenous...of photobleaching, autofluorescence, and, in the case of exciting cyan-excitable donors, phototoxicity...

  2. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...Thakore PI, Majoros WH, Reddy TE, Gersbach CA. Multiplex CRISPR/Cas9-based genome editing for correction of...Ran FA, Yan WX, Asokan A, Zhang F, Duan D, Gersbach CA. In vivo genome editing improves muscle function ...Perez-Pinera P, Brown MT, Majoros WH, Reddy TE, Gersbach CA. Correction of dystrophin expression in cells from...long period of time. High fidelity Cas9s such as eSpCas9 and SpCas9-HF should be explored to reduce off-target...mutation in dystrophin exon 23. Long et al. used SpCas9 in conjunction with AAV9, whereas Nelson et al....al. and Tabebordbar et al. used the shorter SaCas9 with AAV8 and AAV9, respectively. Although each study... it’s important to realize that DMD is a simpler case than other genetic diseases we’d like to treat with...

  3. The 12 Days of CRISPR

    Type
    Blog Post
    ... 11th day of CRISPR, Addgene gave to me: Cas-E-le-ven! (Cas11)🎵 On the 10th day of CRISPR, Addgene gave...🎵 On the 3rd day of CRISPR, Addgene gave to me: dCas9-based gene editing for cleavage-free genomic knock-in...day of CRISPR, Addgene gave to me: High-fidelity Cas13 variants for targeted RNA degradation with minimal...

  4. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nat Biotechnol...Photoswitchable gRNAs for Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation. ACS 6, 5, 695-703 (2020...surprise then that stability of the RNA used in CRISPR-Cas9 experiments – the “gRNA” – suffers from the same...still effectively serving their function of guiding Cas9 to generate a targeted break. These advances in ...modifications The recognition sequence for CRISPR-Cas9 targeting is led by a ~20 bp gRNA which is part ...trans-activating crRNA (tracrRNA) which interacts with the Cas9 protein. These RNA molecules are vulnerable to attack...partially DNA gRNA is surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency. In...

  5. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ... it’s easy to switch out reporters, Cas proteins, guide cassettes, promoters, N’ and/or C’ terminus, and...specific to the type of module it is (Pol 2 protomer, Cas protein, ect...). Whatever individual components ...above selections.) CRISPR enzyme (Cas9 [sp], Cas9 [other], Cas12a, RNA-targeting) CRISPR mechanism ...destination vectors contain an ampicillin resistance cassette and an origin of replication, which are not schematized...using more than six or seven components — in most cases, that's making two or more plasmids — it may be ...

  6. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...Hypercompact genome editor CRISPR-Casɸ By Melina Fan The Doudna Lab adds to the CRISPR-Cas toolbox with their discovery...well-characterized Cas enzymes, which rely on multiple active sites. In addition, Casɸ has a minimal PAM...three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several reasons. Casɸ is only...70kDa, with a molecular weight roughly half of Cas9 or Cas12a. This is in part due to the fact that it only...demonstrate that CRISPR-Casɸ is active in vitro as well as in human and plant cells. Casɸ from Biggiephage...Blue-light inducible Cre for use in bacteria CRISPR-Casɸ mGold, a new yellow fluorescent protein New CRISPR...excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases to manipulate...

  7. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ... dehalogenase that binds a synthetic chloralkane (CA) ligand. The current standard version known as HaloTag...TMP-tag) Few Easy HaloTag 295 Chloralkane (CA) Many Easy CLIP-tag 182 Benzylcytosine...) COS-7 cell ER labeled with Halo-Sec61β with SiR-CA (magenta) and outer mitochondrial membrane labeled...that your fusion protein is functional. In some cases, a small peptide tag like tetracysteine might be...

  8. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...impaired Cas protein (dCas or Cas nickase)  a guide RNA that targets the complex of TadA and dCas to the.... pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG) to...constructs using a catalytically “dead” S. pyogenes Cas9 (dC9-ABE8.x-m/d). The dC9-ABE8.x-m/d constructs ...frequency more than 90% compared to ABE7.10. Choosing Cas9 proteins to increase the targeting scope of adenine... base editors The most commonly used S. pyogenes Cas9 requires the target site in the genomic DNA to contain... to create NG-ABE8.xm/d and the S. aureus Cas9 (PAM: NNGRRT) to create Sa-ABE8.x-m/d. With these variants...frequency over ABE7.10. The ability to use non-standard Cas9 in the ABE8 complexes enables a broader targeting...

  9. CRISPR Cheat Sheet

    Type
    Blog Post
    ...However, prime editing and the identification of other Cas proteins continue to expand the targeting range. ...the gRNAs to cut particular DNA or RNA sequences. Cas9 is a very commonly used CRISPR nuclease. Note:...versions of CRISPR nucleases that can’t cut DNA (dCas9, for example) that are used to increase or decrease...

  10. Hot Plasmids - August 2020

    Type
    Blog Post
    ...The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base ...multiplexed gRNAs and of dCas9 fusions. The dCas-KRAB activator and the dCas-VPR repressor are introduced...an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor in hPSCs is also driven...piggyBac gRNA delivery and the all-in-one dCas9-KRAB and dCas9-VPR targeting plasmids can be found at Addgene...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly... could quantify and track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector ...plasmids for piggyBac gRNA delivery and the all-in-one dCas9 targeting plasmids! Hazelbaker et al., Sci Reports...

  11. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...by our guest bloggers, the experts in the CRISPR/Cas field, have been extremely helpful for scientists...Plasmids 101: Yeast Vectors By Marcy Patrick CRISPR-Cas9 FAQs Answered! By Caroline LaManna Plasmids 101...

  12. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...with Addgene which allow scientists to use CRISPR-Cas technology to perform genome-wide knockout screens...these new CRISPR screening tools, see our CRISPR/Cas Plasmids: Pooled Libraries webpage or read our blog...Knockout Screening. Also, check out our updated CRISPR-Cas resources at www.addgene.org/CRISPR/ to browse CRISPR...information on CRISPR technology? See our improved CRISPR-Cas Guide. Wang et al., Science. 2014. Jan 3;343(6166...Guilinger et al., Nat Biotechnol. 2014..) Our CRISPR-Cas collection of plasmids updates frequently, so visit...plasmids available! Interested in purifying Cas9? Check out pET-28b-Cas9-His from Alex Schier's lab, designed...the lab of Yonglong Chen, pCS2-3xFLAG-NLS-SpCas9-NLS is a Cas9 expression plasmid that was used by Chen...

  13. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...For this, some online tools such as Cas-OFFinder will suffice. Cas-OFFinder provides a bare-bones data...prove somewhat useful for the latest NTT targeting Cas protein, Cpf1. Separately, on-target efficacy is ... tools yet. As a genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of...only one, or a subset of organisms. In only a few cases the software is organism agnostic. Most tools are...will find that some tools cater to specific use-cases through the way their algorithms were developed ...with R. For the Java enthusiasts, take a look at sgRNAcas9. Top tools for transferring CRISPR technology ... parasite Plasmodium falciparum using the CRISPR-Cas9 system." Nature biotechnology(2014). PubMed PMID...

  14. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...-wide profiling of off-target cleavage by CRISPR-Cas nucleases." Nature biotechnology 33.2 (2015): 187...PMC4320685. 3. Fu, Yanfang, et al. "Improving CRISPR-Cas nuclease specificity using truncated guide RNAs."...effects of gRNA and Cas9 expression, they fail to account for the effects of Cas9-induced dsDNA breaks... either stably expressed Cas9 or that had been lentiviral infected with Cas9. Validating the Library The...Genome-wide CRISPR/Cas9 screens are a high-throughput systematic approach for identifying genes involved.... Unanswered questions about genome-wide CRISPR/Cas9 screen design Are non-targeting guides an appropriate...single-guide studies? Many strategies exist for reducing Cas9 off-target effects, but most studies have focused...

  15. Fueled by Coffee at #SfN14

    Type
    Blog Post
    ...I’ve seen evidence of the infiltration of CRISPR-Cas tools into neuroscience experiments. Speaking of...

  16. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...Proteins Switch Off CRISPR-Cas Systems.  Listen to the anti-CRISPR podcast segment Simply expressing anti-CRISPR... group of proteins that are able to block CRISPR-Cas systems. They have been used to block genome editing....2014.12.011 Polstein LR, Gersbach CA (2015) A light-inducible CRISPR-Cas9 system for control of endogenous...applied to create photoswitchable spCas9, dCas9, and S. aureus Cas9 (saCas9) proteins, showing the versatility...specificity as full-length Cas9 (flCas9). When triggered by blue light (470nm), paCas9-1 induced indel mutations...proteins from N. crassa Split Cas9, Split Cas9 nickase, Split dCas9 Genome editing by NHEJ and HDR, repression...Photoswitchable Cas9 Richter et al., 2016 tsRC9 RsLOV from R. sphaeroides Cas9, dCas9 Gene editing ...

  19. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...with inactive Cas enzymes can alter gene expression via acetylation or demethylation. Casilio DNA Methylation...fluorescent dCas9-KRAB and dCas9-VPR for quantifying and tracking cells that express both the dCas9 construct... lower average Cas9-independent off-target DNA editing, ~5–50-fold lower levels of Cas9-dependent off-...Caulobacter genes using a catalytically dead Cas9 (dCas9) derived from Streptococcus pasteurianus  or ...Saccharomyces cerevisiae. The system consists of dCas9, the mammalian transcriptional repressor Mxi1, and...accessible and the complex enters the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the...s lab identified and validated CBEs with reduced Cas9-independent deamination. They generated these editors...

  20. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ...Plasmids CRISPR Information at Addgene More CRISPR-Cas Blog Posts    ...Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9" from the Journal of Visualized Experiments (JOVE...and Daniel Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines...motivations behind this research.   Using CRISPR/Cas9 for targeted genomic deletions We were inspired ...with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore the paired double-strand break...disrupted cells can be easily identified. Even in the case of clones with monoallelic deletion, the nondeleted...regularly interspaced palindromic repeats (CRISPR)/Cas9 nuclease system in mammalian cells. J.Biol.Chem....
Showing: 141 - 160 of 714 results