...1, 2, 5, 9 Deisseroth 26973 pAAV-hSyn-hChR2(H134R)-EYFP Syn ChR2/H134R EYFP Constitutive 1, 2, 5, 9, rg...Constitutive 1, 5, rg* Boyden 59171 pAAV-Syn-ChrimsonR-tdT Syn ChrimsonR tdTomato Constitutive 1, 5, 9 Boyden... dependent 5 Boyden 99039 pAAV-CamKII-ArchT-GFP (PV2527) CaMKII ArchT GFP Constitutive 1, 5, 9, rg* Boyden...Constitutive 5, 8, rg* Boyden 84445 pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] CAG Jaws GFP Cre dependent 1, 5, rg* ...Constitutive 1, 5, rg* Yizhar 125713 AAV-hSyn1-SIO-eOPN3-mScarlet-WPRE Syn eOPN3 mScarlet Cre dependent 1, 5 Yizhar...Constitutive 1, 5 Yizhar 198511 pAAV_hSyn-DIO-PdCO-mScarlet-WPRE Syn PdCO mScarlet Cre dependent 1, 5 Yizhar 198513... 1, 5 Yizhar 198516 pAAV_EF1a-DIO-PdCO-mScarlet-ER-miniWPRE EF1a PdCO mScarlet Cre dependent 1, 5 Yizhar...

...(A & B) Knockout Mouse X.S. Liu 3rd 5 5 10 49,252 Human Metabolic Gene CRISPRa sgRNA Library 187080 Activation...Weissman 3rd 5 10 104,540 209,080 CRISPRa-v2 83996 1000000093 Activation Mouse Weissman 3rd 5 10 107,105...Weissman 3rd 5 10 104,535 209,070 CRISPRi-v2 83987 1000000092 Inhibition Mouse Weissman 3rd 5 10 107,415...Weissman 3rd 5 Varies hCRISPRi-v2 subpooled libraries 83971 — 83977 Inhibition Human Weissman 3rd 5 Varies ...Weissman 3rd 5 Varies mCRISPRi-v2 subpooled libraries 83989 — 83995 Inhibition Mouse Weissman 3rd 5 Varies ...Mouse Yusa 3rd 5 87,897 Mouse improved genome-wide library v2 67988 Knockout Mouse Yusa 3rd 5 90,230 Mouse... Library v2 131625 Inhibition E. coli Bikard N/A ∼5 21,417 Bison sgRNA Library 169942 Knockout Human Ebert...

...Cre-dependent 5, 9 Sternson 119742 AAV SYN PSAM4 GlyR IRES EGFP PSAM4 GlyR - Inhibition IRES EGFP none 5 Sternson...EGFP none 5 Sternson 121538 pAAV SYN1 HA-hM4D(Gi) hM4D(Gi) - Inhibition HA fusion none 2, 5 Richie 121539...mCitrine none 5 Roth 50478 pAAV-GFAP-hM3D(Gq)-mCherry hM3D(Gq) - Activation mCherry fusion none 5 Roth 50479...) - Activation mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg*, PHPeB Roth 44362 pAAV-hSyn-DIO-hM4D(Gi...) - Inhibition mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg*, PHPeB Roth 65417 pAAV-hSyn-dF-HA-KORD-...mCherry hM3D(Gq) - Activation mCherry fusion none 1, 2, 5, 8, 9, rg* Roth 50475 pAAV-hSyn-hM4D(Gi)-mCherry hM4D... hM4D(Gi) - Inhibition mCherry fusion none 1, 2, 5, 8, 9, rg* Roth 52536 rAAV-CAG::FLEX-rev:: hM4D-2a-...

...Cre.SV40 CamKII none 1, 5, 9, rg* Wilson 105537 pENN.AAV.CMVs.Pl.Cre.rBG CMV none 1, 2, 5, 8, 9, rh10 Wilson...pAAV.GFAP.Cre.WPRE.hGH GFAP none 5, PHPeB Wilson 196410 AAV-GfaABC1D-Cre-4x6T GfaABC1D none 5 Carmichael 107788 AAV.rTH.PI.Cre.SV40...EBFP-Cre Syn EBFP 5 Zeng 105540 pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 Syn eGFP 1, 2, 5, 8, 9, rg*, PHPeB...105545 pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 CMV eGFP 1, 2, 5, 8, 9 Wilson 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a...Deisseroth 121675 pAAV-EF1a-fDIO-Cre EF1a none 1, 5, 8, 9, rg* Engel , Nectow 69570 pAAV-EF1a-N-CretrcintG... 105553 pENN.AAV.hSyn.Cre.WPRE.hGH Syn none 1, 2, 5, 8, 9, rg* Wilson 105555 pENN.AAV.hSyn.Cre.hGH Syn...107738 pAAV-hSyn-Cre-P2A-dTomato Syn dTomato 1, 2, 5, 8, 9, rg*, PHPeB Larsen 107787 AAV.TBG.PI.Cre.rBGe...

... HTR3A 5-hydroxytryptamine (serotonin) receptor 3A 5-HT-3, 5-HT3A, 5-HT3R, 5HT3R, HTR3 HTR3B 5-hydroxytryptamine...heavy diversity 5-24 (non-functional) IGHD524 IGHD5-5 immunoglobulin heavy diversity 5-5 DK4, IGHD55 IGHD6...:14099, K5-5, MGC88293 CCR5 chemokine (C-C motif) receptor 5 CC-CKR-5, CCCKR5, CD195, CKR-5, CKR5, CMKBR5...receptor 3B 5-HT3B HTR3C 5-hydroxytryptamine (serotonin) receptor 3, family member C - HTR3D 5-hydroxytryptamine... interleukin 5 (colony-stimulating factor, eosinophil) EDF, IL-5, TRF IL5RA interleukin 5 receptor, alpha...

... four Venus fluorescent proteins connected via a 5, 5 and 6 amino acid linker, respectively VVVVV Anisotropy...Plasmid FRET Pair C5V Cerulean attached to Venus via a 5 amino acid linker C17V Cerulean attached to Venus ... acid linker C5A Cerulean attached to Amber via a 5 amino acid linker C17A Cerulean attached to Amber ...of two Venus fluorescent proteins connected via a 5 amino acid linker V17V Anisotropy/brightness standard... three Venus fluorescent proteins connected via a 5 and 6 amino acid linker, respectively VVVV Anisotropy... of five Venus fluorescent proteins connected via 5 amino acid linkers VVVVVV Anisotropy/brightness standard...consisting of six Venus fluorescent proteins connected via 5 amino acid linkers pET28CLY1 Peptide linker standard...

...Description PI 16539 pBV-Luc Firefly Insertion of 5' promoter/enhancer regions Bert Vogelstein 12178 pIS0...normalization William Kaelin 71248 pXPG Firefly Insertion of 5' promoter/enhancer regions. Mutations within the luciferase...Cockerill 64784 pGL3-Basic-IRES Firefly Insertion of 5' promoter/enhancer regions Joshua Mendell 212936 pGL3... Groskreutz 60323 pGL4.23-GW Firefly Insertion of 5' promoter/enhancer regions using Gateway cloning. ...luciferase and renilla_polyA Firefly/Renilla Insertion of 5' promoter/enhancer regions upstream of Firefly luciferase.... Mikko Taipale 87789 pPTluc Firefly Insertion of 5' promoter/enchancer regions in Drosopholia Herman ... Calabrese 106253 pLS-mP-Luc Firefly Insertion of 5' promoter/enhancer regions. Lentiviral Nadav Ahituv...

...parameters: 37 °C for 30 min; 95 °C for 5 min and then ramp down to 25 °C at 5 °C/min. Dilute oligos 1:10 in ddH...following parameters: Cycles 1-20 (37 °C for 5 min, 20 °C for 5 min); Cycle 21 (80 °C for 20 min). These ...enzyme (5,000 U/ml), 5.0 μl ATP (10 mM), 0.25 μl (5 μg) BSA (20 mg/ml), 0.375 μl (750 U) T4 DNA ligase... solution and add to electroporation cuvette. Add 5 μg of each CRISPR/Cas9 construct (10 μg total). Add...construct. Electroporate cells with 250 volts for 5 msec in a 2 mm cuvette using an electroporation system...population exposed to sgRNA-A and sgRNA-B (see step 5 ). The purpose of the following PCR is to validate...multichannel pipette. Centrifuge PCR plate at 400 x g for 5 min and remove supernatant by flicking the PCR plate...

...bacterium (LbCpf1) 5' TTTV AsCpf1 RR variant 5' TYCV LbCpf1 RR variant 5' TYCV AsCpf1 RVR variant 5' TATV Campylobacter...sequence may not be characterized *In the table above, 3' and 5' indicate on which end of targeted sequence...will continue to anneal to the target DNA in a 3′ to 5′ direction. The location of any potential mismatches...inhibit target cleavage, while mismatches toward the 5′ end distal to the PAM often permit target cleavage...from naturally occurring cytidine deaminases. Figure 5: Overview of base editing More recently, researchers...transcription. The edited DNA strand displaces the unedited 5’ flap, and the resulting heteroduplex is resolved ... repair using the edited strand as a template PE4/5 - improved PE2/PE3 to inhibit mismatch repair to promote...

... actual oligo you should order is: 5'-CACCGN...N-3' 3'-CN...NCAAA-5' You need to add the CACC and AAAC...purification and have tried conditions with 5% DMSO or without 5% DMSO. My genomic PCR didn’t work, do you... http://www.cell.com/abstract/S0092-8674(13)01015-5 As for target selection, we usually pick 3-6 guides... Feng Zhang's Lab, has assembled a list of FAQs about using the lab's CRISPR technology. Genome..., February 2013 ), has assembled a list of FAQs about using the lab's CRISPR technology. Visit the Zhang...

...SF-1, SF1, SPGF8, SRXY3 nuclear receptor subfamily 5, group A, member 1 NR5A2 Plasmids NR5A2, B1F, B1F2...FTZ-F1beta, LRH-1, LRH1, hB1F-2 nuclear receptor subfamily 5, group A, member 2 NR6A1 Plasmids NR6A1, GCNF, GCNF1...nuclear receptor plasmid collection assembled in collaboration with the Nuclear Receptor Signaling Atlas (NURSA...associated with numerous disease pathways. In collaboration with research labs from around the globe, Addgene...

...and breathing, and often results in death within 3-5 years from symptom onset. Disease mechanisms are still...having no prior family history. A small percentage (5-10%) are familial ALS cases having at least one other...knockout . Maimaitili et al. Nat Commun. 2023 Dec 5. See More Antibodies Find information on Addgene's...Find out about the plasmid collections available at Addgene that may be useful for neurodegeneration ...Scientists are studying protein aggregation, learning about the blood brain barrier integrity, and using CRISPR...with support from the BRAIN Initiative. Jackson Laboratory (Link opens in a new window) A collection of ...

...Event Details Date: Friday September 21, 2018, 9 AM - 5 PM Location: Addgene, 75 Sidney St, Cambridge, MA,...00 PM Wrap Up & Next Steps Melina Fan/Leila Haery 5:00 PM Adjourn Meeting Preparation For Speakers: Each...Learn about Addgene's 2018 AAV Education and Development (AAVED) workshop series. The... Institute Katherine Matho Cold Spring Harbor Laboratory Gowan Tervo Janelia Research Campus Sarada Viswanathan...Institute Katherine Matho, Cold Spring Harbor Laboratory Gowan Tervo and Sarada Viswanathan, Janelia Research...

...Cell Chem Biol. 2016 Jun 1. pii: S2451-9456(16)30163-5. Teresa Alonso , Javier García-Sancho Calcium Genetically... calmodulin-peptide pairs. Chem Biol. 2006 May;13(5):521-30. Amy Palmer , Roger Tsien Calcium Turquoise... in the endoplasmic reticulum. Biophys Rep (2019) 5, 31–42 Pingyong Xu Magnesium Fluorescent Mg2+ sensor...influx/kynurenine efflux cycle. PLoS Biol. 2007 Oct;5(10):e257. Wolf Frommer Return to Top Neurotransmitters...2023.05.27.542566. Yulong Li Serotonin Serotonin (5-HT) sensor GRAB_5-HT1.0 for viral expression in neurons...measuring serotonin dynamics. Nat Neurosci. 2021 Apr 5. Yulong Li , J. Julius Zhu Serotonin Serotonin sensor...S0092-8674(20)31612-3. Lin Tian Serotonin Serotonin (5-HT) sensor psychLight Psychedelic-inspired drug discovery...

...AAV5, AAV8, or AAVrg) and volume (1 mL, 2.5 mL, or 5 mL). Currently, we can only package plasmids in our...100 µL aliquots 2.5 mL Large 50 × 100 µL aliquots 5 mL *AAV2 yields are generally lower, with a minimum...Additional Resources Viral Vector Production Learn about how we produce high-quality viral vector preps at...

...(or Cas9 nickase) Single (or dual) gRNA targeting 5′ exon or essential protein domains High-fidelity Cas...interest. For genetic knockouts, gRNAs commonly target 5′ constitutively expressed exons, which reduces the...CRISPR/Cas9-based genome editing. Scientific Reports . 5 , 11315. PMID: 26066060 Zuris, J. A., Thompson, D....components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable, tunable...when carrying out CRISPR experiments. Read more about how to design your gRNA . Browse Plasmids: gRNAs...to optimize your delivery conditions. Learn more about CRISPR delivery in mammalian systems . Validate ...

...slight melting of the 5 ́-terminus during the reaction. This “peeling back” of the 5 ́-phosphorylated base...anyway? Let’s talk about it. DNA ligases are enzymes that seal breaks in DNA by joining 5 ́-phosphorylated... in vitro, Taq DNA ligase will ligate only nicks (5-8). Taq ligase is a NAD+-dependent DNA ligase from... the downstream side (the base pair providing the 5 ́-phosphate to the ligation). The structural/mechanistic...PMID: 19329793. PubMed Central PMCID: PMC2719376. 5. Barany, Francis. "Genetic disease detection and DNA...Veronique Bailly, and Walter G. Verly. "Nicks 3′ or 5′ to AP sites or to mispaired bases, and one-nucleotide...reaction in a PCR world." PCR Methods Appl 1.1 (1991): 5-16. PubMed PMID: 1842922. 13. Wiedmann, M., et al....

...Kyprianou K. World J Gastrointest Oncol. 2013 May 15; 5(5): 97–101. doi: 10.4251/wjgo.v5.i5.97. PubMed PMID... Frederick National Laboratory for Cancer Research . Frederick National Laboratory for Cancer Research...

...12 (5 min., 37 ºC -> 5 min., 16 ºC) x 30 (5 μl) 245 99.5% 48,900 489,000 24 (5 min., 37 ºC -> 5 min....amplicon form (shown above) are flanked by BsaI sites in an orientation that results in 5’ four base overhangs...similar Type IIS restriction enzyme cutting results in 5’ four base overhangs complementary to the overhangs... and accurate single-insert assembly Intermediate 5- or 12-fragment assembly More complex 24-fragment ...frequencies as the antibiotic selection markers. The 5-, 12- and 24-assembly systems were based on the ability...pGGA destination plasmids. A final validation of the 5-, 12- or 24-fragment test systems was performed by...assemblies (Figure 3, Table 1). These improvements (5- to 12-fold increases in the number of transformants...

...restriction site (GAATTC) to the 5’ end of this primer, making our Forward Primer 5'-GAATTCATGTGGCATATCTCGAAGTAC...consist of: Leader Sequence: Extra base pairs on the 5' end of the primer assist with restriction enzyme ...Therefore, our Forward Primer will use the sequence 5'-ATGTGGCATATCTCGAAGTAC-3' for the region that binds..., resulting in a final Forward Primer sequence of 5'-TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse...final 18 bases of the ORF, including the stop codon (5'-TGGCATATCTCGAAGTACTGA-3'), then adding NotI (GCGGCCGC...restriction enzyme digestion. This gives us a sequence of 5'-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3' (30bp with...put the sequence we chose for our reverse primer (5’-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this...