...donor DNA design, and implemented our optimised protocols into a medium-throughput pipeline for epitope ... mammalian cell types. A detailed step by step protocol for the csRNP method is described in our recent...

...passion for photosynthetic communities, microbial interactions, and science education. Photo cred Alina...

... cerevisiae - Dueber Lab Yeast Prototrophy : Restoration of prototrophy in common S. cerevisiae lab strains...microbe of interest, including bacteria, viruses, protozoa, fungi, and more. Plasmid...including bacteria, viruses, parasites such as protozoa, and fungi. Find plasmids below for the species...Cryptococcus sp. Plasmids for Prions Prions Plasmids for Protozoa Species Entamoeba histolytica Leishmania sp. Plasmodium... page for CRISPR resources, such as depositor protocols and gRNA design software. Addgene yeast CRISPR...

...contains many unique protospacer sequences that have homology to foreign DNA. Protospacers are separated by...Engineering You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR...contain a species-specific sequence known as a protospacer adjacent motif (PAM). The CRISPR complex binds...sequence is only valid if it contains a special Protospacer Adjacent Motif (PAM) directly after where the...target RNA rather than DNA, sometimes requiring a protospacer flanking sequence (PFS). In bacteria, Cas13 targeting...further develop the CRISPR toolkit by posting lab protocols , providing tips from experts in the field, and...

...hydroxytryptamine (serotonin) receptor 3B 5-HT3B HTR3C 5-hydroxytryptamine (serotonin) receptor 3, family...RIFLE, TAL LTA lymphotoxin alpha (TNF superfamily, member 1) LT, TNFB, TNFSF1 LTB lymphotoxin beta (TNF superfamily...member 1 CD94 LGMN legumain AEP, LGMN1, PRSC1 LTA lymphotoxin alpha (TNF superfamily, member 1) LT, TNFB, TNFSF1... RNase A family, 2 (liver, eosinophil-derived neurotoxin) EDN, RNS2 ROBO1 roundabout, axon guidance receptor...3 HIS2, HTN2, HTN5 HTR3A 5-hydroxytryptamine (serotonin) receptor 3A 5-HT-3, 5-HT3A, 5-HT3R, 5HT3R, HTR3...family member C - HTR3D 5-hydroxytryptamine (serotonin) receptor 3 family member D MGC119636, MGC119637 ...MGC119637 HTR3E 5-hydroxytryptamine (serotonin) receptor 3, family member E 5-HT3c1, MGC120035, MGC120036, MGC120037...

...pAAV-CAG-iSeroSnFR-Nlgn Fluorescent reporter for serotonin (dendrite localization tag) Lin Tian 135420-AAV1...parapinopsin driven by the CAG promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades...parapinopsin driven by the Ef1a promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades...frame with EGFP under control of human synapsin1 promotor. Ofer Yizhar 198513-AAV5 pAAV_hSyn-PdCO-EGFP-WPRE...frame with EGFP under control of human synapsin1 promotor. Ofer Yizhar 198516-AAV1 pAAV_EF1a-DIO-PdCO-mScarlet-ER-miniWPRE...optimized PdCO under control of human synapsin1 promotor after Cre-dependent recombination Ofer Yizhar ...

... (2) a complementary DNA sequence, and (3) a protospacer-adjacent motif (PAM) on the opposite strand. ...this strand is edited by base editors! PAM Protospacer-Adjacent Motif A short sequence that must be ...

...important to choose a target sequence with a nearby protospacer-adjacent motif (PAM)? Cas9 is becoming an ever...10.1016/j.cell.2014.02.001 Osuka, S., Isomura, K., Kajimoto, S., Komori, T., Nishimasu, H., Shima, T., Nureki...

...) CRISPR software and depositor protocols JOVE video reprint: Protocol for genomic deletions in mammalian...target is present immediately adjacent to a P rotospacer A djacent M otif (PAM) The PAM sequence (NGG)...can increase the risk of off-target cutting or cytotoxicity, but naturally-occurring Acr (Anti-CRISPR) proteins...fusing the anti-CRISPR protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled...before use (Figure 8C), following the amplification protocol specified by the depositing lab. After amplification...The translated codons that make up a gene PAM P rotospacer A djacent M otif; Sequence adjacent to the target...engineering using the CRISPR-Cas9 system. Nature Protocols , 8 (11), 2281–2308. PMID: 24157548 Ran, F. A....

...but the good news is that you only need one. ProtospacerWB was developed with the purpose of applying CRISPR..., online), DESKGEN (commercial, online), or ProtospacerWB (academic, offline). While it remains to be ...

...Additional Resources: CRISPR protocols from Addgene depositors JOVE Video Reprint: Protocol for genomic deletions...review the protocol associated with the specific plasmid in question (see CRISPR protocols from Addgene...Biology . 1239 , 197–217. PMID: 25408407 Hashimoto, M., & Takemoto, T. (2015). Electroporation enables the...

... Large Stokes Shift , Photoactivatable , Photoconvertible , and Photoswitchable fluorescent proteins Various...monitor in live cells (beware of artifacts!), photoactivatable tags for super resolution imaging, epitope...20341 mEos2 (fluorescent protein) Loren Looger Photoactivable Fluorescent Protein Expression: contains mEos2...mEos2 with an N-terminal His tag. Photoactivable fluorescent proteins like this one can be used in super...

..., ABE8s resulted in 1.5-fold more editing at protospacer positions A5-A7 and 3.2-fold more editing at ...Nishida, K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki, M., Miyabe, A., Araki...

...upper and lower motor neurons in the brain and spinal cord. Over time, a loss of motor neurons leads to...aberrant axon initial segment (AIS) plasticity with a motor neuron-specific reporter and a PAX7 inducible vector...expression of transcription factors for sensory, motor, and cortical neuron differentiation. Held et al...opens in a new window) Research workspace on Protocols.io (Link opens in a new window) . This resource...

...Near-Infrared Long Stokes Shift Photoactivatable Photoconvertible Photoswitchable Timers Blue/UV Protein Excitation...LSSmKate2-C1 - Mammalian Expression Jump to Top Photoactivatable (e.g. off to on) Excitation and Emission wavelengths...HisB-PAiRFP2 - Bacterial Expression Jump to Top Photoconvertible (e.g. green to red) Protein Excitation (nm...pBAD-PSmOrange2 - Bacterial Expression Jump to Top Photoswitchable (e.g. off to on to off) Protein Excitation ...

...pAAV-CIBN-CreC photo-activatable cre recombinase CAG AAV Bruchas 75268 pAAV-CRY2-CreN photo-activatable ...Cepko 13780 pNrl-Cre Cre-Myc, Expressed in rod photorecetor cells in the retina Nrl Mammalian Cepko 13962...pAAV-Syn-iCre iCre AAV Ritola 122960 pcDNA3.1_PA-Cre photo-activatable Cre recombinase CMV Mammalian Sato 122961...pBbE5a-Opto-Cre-Vvd Cre recombinase split with Vivid photodimers lacUV5 Bacterial Dunlop 135217 pDEST mfap4:icre-p2a-tomato...pAAV-EF1a-NCreV N-terminal Cre component split with Vivid photodimers EF-1 alpha AAV Zeng 140132 pAAV-EF1a-CCreV C-...C-terminal Cre component split with Vivid photodimers EF-1 alpha AAV Zeng 140135 pAAV-EF1a-iCreV Split...

...Information Cloning Protocols Single guide cloning: Zhang Lab General Cloning Protocol (PDF, 237 KB) Dual...Dual guide cloning: Brzezinski Lab Dual Guide Protocol (PDF, 990 KB) Plasmid and Sequence Derivative Sources...Prdm1 and Vsx2 enhancers in the retina alters photoreceptor and bipolar cell fate specification, yet differs...

... researchers face right now (e.g., not enough protocols, cost, not enough good antibodies, access to reagents...practical information in the form of data and protocols showing examples of AAV strategies for intersectional...or any other useful information. Experimental protocols or schematics. Examples of research materials ...Retrograde hSyn1-GCaMP6f-P2A-nls-dTomato Detailed protocol: AAV Production in HEK293 Cells Please share materials...

... #2 Library (NEDD4-2 WW2 domain) References & Protocols Iterative Synthetically Phosphorylated Isomers....202110591. PubMed (Link opens in a new window) Protocols for using iSPI, rEcoli XPS, and pSerOTS-C1* (V70....2015.08.016. PubMed (Link opens in a new window) Protocol for using the Improved Reagents (ca. 2015) from...):3716-22. PubMed (Link opens in a new window) Protocol for Reagents (ca. 2014) from the Rinehart lab:...

... 1000000030 Yamamoto Lab TALEN Construction and Evaluation Accessory Pack Takashi Yamamoto This accessory...Golden Gate. Validated in several species. Takashi Yamamoto Additional Plasmids for Use with Addgene TALEN... at the final assembly step in the Golden Gate protocol. Both plasmids 47388 and 47389 include a Flag ...