...a zinc finger (ZF) nuclease. Zinc fingers recognize triplets and the FokI nuclease operates as a dimer...sites. (B) A TAL effector nuclease (TALEN). Similar in principle to the ZF nuclease, the components of the...these tools and the researchers who are learning to wield them. Learn more about Addgene’s TAL effector and...years of research has shown that when fused to a nuclease or activation domain, a ZF array is an effective...at Massachusetts General Hospital also recently released a TALEN kit through Addgene. This kit, first described...

...amplify. Typically use primers at least 100 bp from the predicted cleavage site to ensure detection would...322.2 KB If you have trouble loading this video, please try viewing on YouTube . Abstract The prokaryotic...lines. Protocol For referenced Tables and Figures, please access the manuscript PDF posted at the top of ...genomic matches or near-matches to reduce risk of cleavage away from target sites (off-target effects). Ensure...which are likely to cause frameshift mutations leading to nonsense mediated decay of the mRNA transcript...of primers upstream and downstream of the sgRNA cleavage sites (“deletion band”; Figures 1 - 2 ). In the...for scarring (small indels produced at the sgRNA cleavage site without the intended deletion). Use a pair...

...code inserted into the plasmid. This produces the least amount of variation between antibodies, allows for... induced to express antibodies. Antibodies are released into the growth medium and can either be used ...fragments - Fab fragments are created by enzymatically cleaving the variable regions of an antibody (each Fab ...they should be stored at -20 °C in aliquots of at least 10 µL. Antibodies cannot tolerate freeze-thaw cycles...single cells. Includes: Flow cytometry Read on to learn more about the applications common to each category...should be run with your protein of interest and at least one ubiquitously expressed protein such as GAPDH...protein:DNA complexes. After capture, the DNA is released from the complex and can be sequenced and amplified...

...MDH). The MCC is a national group of educational leaders providing research opportunities to their students...MDH and, if indicated on the plasmid map, a TEV cleavage site is present between the 6xHis-tag and the ... region of MDH. For detailed clone information, please see the MDH Protein Clone Information Sheet, available... CUREs Community (Link opens in a new window) — Learn how to get started with a CURE, explore MDH projects...projects, find teaching resources and protocols, learn about regional hubs, access a teaching mentor, and...window) External Collaboration Results in Student Learning Gains and Positive STEM Attitudes in CUREs. Callahan...undergraduate research experiences (CURE) impacts student learning and attitudinal outcomes: A study of the Malate...

... 154948 CaMKII-somBiPOLES-mCerulean CamKII BiPOLES (soma-targeted) mCerulean Constitutive 5, 9, rg* Wiegert...154951 hSyn-DIO-somBiPOLES-mCerulean Syn BiPOLES (soma-targeted) mCerulean Cre dependent 5, 9, rg* Wiegert...Looking For? Please let us know what you would like to see available in viral format. Please note this does...Constitutive Serotype AAV1 AAV2 AAV5 AAV8 AAV9 AAVrg PHPeB Clear Filters ID Name Promoter Opsin Variant Tag Activity...available items found to match all requested criteria. Please consider using fewer filtering options. Browse ...

...CaMKII-somBiPOLES-mCerulean CamKII Bidirectional Optogenetics Wiegert 154951 hSyn-DIO-somBiPOLES-mCerulean Syn Bidirectional...Looking For? Please let us know what you would like to see available in viral format. Please note this does...DREADDs Activators Inhibitors Other Molecular Tools Clear Filters ID Name Promoter Activity Category PI Controls...GCaMP6f-P2A-nls-dTomato Syn GCaMP6f and physically separate nuclear dTomato Calcium sensor Ting 51083 AAV-EF1a-DIO-...AAV-EF1a-DIO-GCaMP6f-P2A-nls-dTomato EF1a GCaMP6f and physically separate nuclear dTomato, Cre-dependent Calcium sensor Ting 83899...GCaMP6s-P2A-nls-dTomato Syn GCaMP6s and physically separate nuclear dTomato Calcium sensor Ting 100853 pAAV.Syn.Flex.NES-jRGECO1a.WPRE.SV40...

...-mediated nucleic acid cleavage. We have harnessed the type II CRISPR nuclease system to facilitate genome...nterspaced S hort P alindromic R epeats) is a microbial nuclease system involved in defense against invading phages...Nature Protocols 2013). For cloning information, please view: Zhang Lab General Cloning Protocol 237.2 ...by co-expressing the S. pyogenes Cas9 (SpCas9) nuclease along with the guide RNA. For most applications...gain-of-function screening. For more information, please visit the Addgene SAM library page . Targeted gene... directed repair-mediated Kras G12D mutations, leading to macroscopic tumors of adenocarcinoma pathology... DNA targeting specificity of RNA-guided Cas9 nucleases. Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann...

...Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. The robust signal, ...sensitivity, wide dynamic range, and resistance to photobleaching has made luciferase a common choice in assays...have made it popular with scientists since its release. Luciferase-containing plasmids are commonly used...applications. Browse the Collection Highlights section to learn more. Highlights of Luciferase Tools in our Collection...artificial luciferase desined de novo using deep-learning approaches. Split-Luciferase and Protein-Protein...luciferase. Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016...linker to the circularly permuted Venus fluorophore. Learn more about pHlash sensor in our 28 Hot Plasmid Technologies...

...protein function results in Rett syndrome is not clear, however, the ability to bind to methylated DNA ...ranscriptional R epressor D omain (TRD) the TRD contains a Nuclear receptor Co-Repressor 1/Silencing Mediator of Retinoic...receptor (NCoR/SMRT) interacting domain (NID) a nuclear localization signal (not essential for localization...RSRT cell line collection (186.8 KB) . To order, please email [email protected] The following cell lines are...information on generating iPSCs using plasmids, please see Addgene's Plasmids for Stem Cell Research page...precise editing of mutations in the MECP2 gene. Please refer to Addgene's CRISPR Guide for a general introduction...your plasmids! To get started with your deposit, please click here ....

...are also available Non-toxic to living cells To learn more about this system, visit the New England Biolabs...website. For a comprehensive comparison to GFP, please refer to this table. Technology SNAP-tag SNAP-tag...permitting selective simultaneous labeling in a sample. Please see the note below for information on substrates...pSNAPf-H2B Control Plasmid SNAP-tag Control for nuclear localization 101123 pSNAPf-ADRβ2 Control Plasmid...pCLIPf-H2B Control Plasmid CLIP-tag Control for nuclear localization 101130 pCLIPf-Cox8A Control Plasmid...SNAP and CLIP tag FAQs 252.2 KB ACP and MCP tags Please note: New England Biolabs (NEB) has recently discontinued...

...conformational change in the sensing region of the construct leads to a change in signal of a circularly permuted fluorescent...Boyden Calcium GCaMP6f/s Ca sensors and separate nuclear localized dTomato fluorophore Ting Lab GCaMP6 Plasmids...sensor (MARIO) A Transient Rise in Free Mg(2+) Ions Released from ATP-Mg Hydrolysis Contributes to Mitotic ...Nagai Magnesium MagFRET-1 Mg2+ sensor, based on Cerulean/Citrine MagFRET: The First Genetically Encoded...3;108(18):7351-6. Amy Palmer Zinc Cytosolic or Nuclear zinc-binding FRET biosensor (Zap) New alternately...through membrane lipid peroxidation, cytosolic release of CAVIN1, and downstream regulation of NRF2. bioRxiv...Ultrafast glutamate sensors resolve high-frequency release at Schaffer collateral synapses. Proc Natl Acad...

...technology. Visit the Zhang Lab page and forum to learn more. CRISPR Design FAQs Homologous Recombination...the Cas9 nickase which will together introduce cleavage of the target site with efficiency similar to ... donor. Generally if off-target (non-specific) cleavage is not a very big concern, you could go ahead ...PX335 vector, you need to find two guides for each cleavage site, but this system probably have better specificity...sequence is required for successful targeting and cleavage, changing it to'NGT' or 'NGC', in addition to ...Cas9 and tracrRNA, it will be processed (i.e., cleaved) into a shorter form that contains 20bp of the ...mature tracrRNA), the spacer used is only 20bp. Please refer to Cong et al. , Science 2013 for more details...

...cloning grade DNA, please let us know using the Cloning Grade DNA Suggestion Form . Please note that this .... Cloning Grade DNA is an Addgene Beta service. Learn more about Addgene's Beta Projects and how you can...µl (± 5ng/µl) Buffer 10 mM Tris-Cl, pH 8.5 Price Please log in to your account and check the individual...I find cloning resources or scientific support? Please see our Addgene Protocols for resources you may...

...Microtubules 87422 LMNB1-mEGFP AICSDP-10 mEGFP Lamin B1 Nuclear envelope 87423 TOMM20-mEGFP AICSDP-8 mEGFP Tom20...114403 LMNB1-mTagRFP-T AICSDP-29 mTagRFPT Lamin B1 Nuclear envelope 114404 AAVS1-mEGFP (PGK) AICSDP-36 mEGFP...NUP153-mEGFP AICSDP-50 mEGFP Nucleoporin Nup153 Nuclear pores 114410 FUS-mEGFP AICSDP-64 mEGFP RNA-binding...fibrillar center) 133964 SON-mEGFP AICSDP-82 mEGFP SON Nuclear speckles 159744 DMD-mEGFP AICSDP-55 mEGFP Dystrophin...PCNA-mEGFP AICSDP-116 mEGFP Proliferating cell nuclear antigen (PCNA) DNA replication foci IPSC cell lines...

...downstream of the sequence to be cleaved. This non-specific cleavage is thought to activate programmed...insertions or deletions (indels) in the DNA. Indels often lead to frameshifts, creating loss of function alleles...DNA. In bacteria, once they have recognized and cleaved the target RNA sequence, they adopt an enzymatically...enzymatically active state and can bind and cleave additional RNAs regardless of homology to the crRNA. This...

...Constitutive 9 Wilson 105557 pENN.AAV.CB7.CI.mCerulean.WPRE.RBG CB7 mCerulean Constitutive 1, 9 Wilson 105598 pAAV.GfaABC1D.PI.Lck-GFP.SV40...Looking For? Please let us know what you would like to see available in viral format. Please note this does...PHP.eB PHP.S PHP.V1 CAP-B10 CAP-B22 MaCPNS1 MaCPNS2 Clear Filters ID Name Promoter Fluorophore/Tag Activity...

...each viral lot. To learn which specific QC experiments were performed on your lot, please contact us . Full...medium (OptiPro SFM) containing lentivirus is first cleared by low-speed centrifugation and then by filtration...responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit (RPP30) as a control for...the sample from the DNA tube all the way to the cleared viral preparation pool. Proviral Identity Confirmation...

...NF1 Neurofibromin 1 NFE2L2 Nuclear factor, erythroid 2 like 2 NFKB1 Nuclear factor of kappa light polypeptide...guanine nucleotide releasing factor RASGRP RASGRP1 RASGRP2 RASGRP3 RASGRP4 RAS guanyl releasing protein RASSF...activating pathways for cell growth and potentially leading to cancer. Constitutively active Ras is the most...

...Disease. These diseases persist for many years and lead to a major burden on the patients, their families.... Scientists are studying protein aggregation, learning about the blood brain barrier integrity, and using...spinal cord. Over time, a loss of motor neurons leads to limb weakness, difficulty speaking, swallowing...percentage (5-10%) are familial ALS cases having at least one other affected family member. These familial... individuals develop late-onset AD. There is no clear association between a gene and late-onset AD. It...

...Looking For? Please let us know what you would like to see available in viral format. Please note this does...mice use LY6A to cross the blood-brain barrier, leading to strain-specific tropism (See S3 table (Huang...review performance data of each of these serotypes, please refer to Addgene’s Data Hub and the Caltech CLOVER...using the Caltech serotypes in future publications, please acknowledge Viviana Gradinaru and cite as follows...