...Collections Fluorescent Proteins Michael Davidson Fluorescent Protein Collection Michael Davidson Fluorescent Protein...Subcellular localization Optogenetics The lab of Michael Davidson from Florida State University has contributed...proteins with your gene of interest. Please visit Michael Davidson's Molecular Expressions website for additional...

...by the Michael J. Fox Foundation... Plasmid Collections MJFF Parkinson's Plasmid Resource Michael J. Fox ...Research Tools (Link opens in a new window) The Michael J. Fox Foundation (MJFF) (Link opens in a new window...which includes other areas of this research topic. Michael J. Fox Foundation Plasmids MJFF has created a number...

...Anti-Pan-Nav1 Na+ channel [N97A/31R] Pan-Nav1 Na+ channel Human Mouse IgG2a 114553 Anti-Pan-Nav1 Na+ channel [N230...Anti-Kv2.1 K+ Channel [D3/71R] Kv2.1 K+ Channel Rat Mouse IgG2a 128613 Anti-Kv1.4 K+ Channel [K13/31R] Kv1.4... Kv1.4 K+ Channel Rat Mouse IgG2a 128614 Anti-Kv1.2 K+ Channel [K14/16R] Kv1.2 K+ Channel Human Mouse ...KCNAB2 K+ Channel [K17/70R] Kvbeta2/KCNAB2 K+ Channel Mouse Mouse IgG2a 128616 Anti-Kv1.1 K+ Channel [K20/....1 K+ Channel Rat Mouse IgG2a 128617 Anti-Pan-Kvbeta K+ Channel [K25/73R] Pan-Kvbeta K+ Channel Mouse ...Anti-Nav1.1 Na+ channel [K74/71R] Nav1.1 Na+ channel Rat Mouse IgG2a 128627 Anti-Kv4.3 K+ Channel [K75/41R] ...] Kv4.3 K+ Channel Rat Mouse IgG2a 128628 Anti-Kv2.1 K+ Channel [K89/34R] Kv2.1 K+ Channel Rat Mouse IgG2a...

...GAL1 Huntington's Michael Sherman 1385 pYES2/103Q HTT Flag, GFP GAL1 Huntington's Michael Sherman 8412 pMTH...Friedreich ataxia Michael Ristow 14977 pCINeo-hFX(I154F)-HA FXN HA, AU1 CMV Friedreich ataxia Michael Ristow 14978... ataxia Michael Ristow 14979 pGL3-basic-hFX-promotor(1,3kb) FXN hFXN Friedreich ataxia Michael Ristow ...ataxia Michael Ristow 14981 pGL3-basic-hFX-promotor(2kb, distal) FXN hFXN Friedreich ataxia Michael Ristow...Friedreich ataxia Michael Ristow 14988 pBlue-hFX(I154F)-HA FXN HA, AU1 T7 Friedreich ataxia Michael Ristow 14989...Friedreich ataxia Michael Ristow 14990 pBlue-hFX(T133E)-HA FXN HA, AU1 T7 Friedreich ataxia Michael Ristow 14991...ataxia Michael Ristow 14992 pET28(a)-hFX(I154F)-HA FXN HA, HIS, AU1 T7 Friedreich ataxia Michael Ristow...

...incorporation Michael Jewett 69495 rEc.13.delA 13 amber sites changed to TAA, RF1 function removed Michael Jewett...Methanogenic archaeon Bacterial Ahmed Badran 209739 pAB228i24 G1 PylRS Methanogenic archaeon Bacterial Ahmed...TAG sites changes to UAG, RF1 function removed George Church 48999 C321 all TAG sites changes to UAG George...sites changes to UAG, RF1 function removed Farren Isaacs 87359 C321.∆A.opt all TAG sites changes to UAG...the tRNA is incorporated into the growing peptide chain. To expand the genetic code, modified tRNAs, codons...amber codon) that is recognized by the cognate charged tRNA. Once these plasmids have been introduced ...machinery. To expand the genetic code, 4 major changes to the standard translation machinery are needed...

...mEmerald* Michael Davidson 54491 mCherry-Lifeact-7 Actin Filaments LifeAct mCherry* Michael Davidson 54610...LifeAct EGFP Michael Davidson 55148 mCherry-Tubulin-C-18 Microtubules alpha-tubulin mCherry* Michael Davidson...mApple Michael Davidson 55165 mCherry-ZO1-C-14 Tight Junctions Zonula Occludens-1 mCherry Michael Davidson...pEGFP-GFP11-Clathrin light chain Clathrin-coated vesicles Clathrin light chain GFP Bo Huang 54040 mEmerald-Clathrin...endosomes RAB7 GFP Richard Pagano 12661 DsRed-rab7 WT Late endosomes RAB7 DsRed Richard Pagano 158006 pCMV-mGold-Lysosome-N...endosomes Rab11a GFP Richard Pagano 12679 DsRed-rab11 WT Recycling endosomes Rab11a DsRed Richard Pagano *Fusions...targeted to the same structures as these well-characterized proteins. General Structures The cell contains...

.... You can therefore change the genomic target of the Cas enzyme by simply changing the target sequence...surface-exposed positively-charged grooves on Cas9. gRNA binding induces a conformational change in Cas9, shifting...Fanton, A., Tycko, J., Hinks, M., Chandrasekaran, S. S., Perry, N. T., Schaepe, J., Du, P. P., Lotfy, P., Bassik...approach. Figure 1: Overview of the basic CRISPR mechanism Engineered CRISPR systems contain two components... In this section, we will introduce the basic mechanisms of CRISPR and some of the ways the CRISPR components...efficiency, or flexibility of the Cas enzyme. Basic Mechanisms of CRISPR The requirements for a CRISPR knockout...target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only cleave a given locus if the gRNA...

..., a conformational change in the sensing region of the construct leads to a change in signal of a circularly... of membrane charges A FRET sensor enables quantitative measurements of membrane charges in live cells...affinities or negative-control constructs used to characterize the sensor's behavior in the target system. ...sensor behavior may differ from the original characterization (e.g. in vitro vs. in vivo , cytoplasmic vs...Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP...GECI) that reports with lifetime and intensity changes A turquoise fluorescence lifetime-based biosensor...calcium sensors (fRCaMP1/2, GECO1/2) The kinetic mechanisms of fast-decay red-fluorescent genetically encoded...

...made up of four protein chains: two large heavy chains and two small light chains. These four domains come... producing antibodies, can change the constant region, and therefore change the isotype of antibodies ...: scFv - Single-chain fragment variables (scFvs) contain only the variable heavy chain and variable light... it is determined by a region called the “heavy chain constant region” or Fc. Different isotypes play ...light chain regions of an antibody. They are small enough to be produced in bacteria (~25 kD), but tend... derived from a camelid antibody called a heavy chain antibody (HCAb). Their small size allows them to... conjugation reaction. Many antibodies can be purchased already conjugated, which can save significant...

...construct expressing mCherry-tagged channelrhodopsin-2 driven by chandelier cell-targeting enhancer Gordon ...213830-PHPeB pAAV_BiCHATe27_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin-2 driven...mCherry AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by SST interneuron-targeting enhancer...mCherry AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by PV+ basket cell-targeting enhancer...dTomato_nlsdTomato AAV construct expressing dTomato driven by chandelier cell-targeting enhancer Gordon Fishell Viviana...mCherry AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by Lamp5 interneuron-targeting enhancer...mCherry AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by VIP interneuron-targeting enhancer...

...promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades. Michael Bruchas 139505-AAV8...promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades. Michael Bruchas 139505-AAV9...promoter. Used for photoswitchable control of inhibitory GPCR signaling cascades. Michael Bruchas 140135-AAV1...AAV9 pAAV-hSyn-DIO-ChrimsonR-mRuby2-ST Cation channelrhodopsin ChrimsonR fused to mRuby2 fluorophore and ...AAV9 pAAV-hSynapsin- soCoChR-GFP A somatic channelrhodopsin, which enables single cell, single millisecond... pAAV-hSynapsin-FLEX-soCoChR-GFP A somatic channelrhodopsin, which enables single cell, single millisecond...pAAV-CAG-DIO-ChroME-ST-P2A-H2B-mRuby3 Cation channelrhodopsin ChroME targeted to the neuronal soma and proximal...

...-strand break. CRISPR systems, found in 95% of archaeal and 48% of bacterial genomes, are highly diverse...defines 5 types and 16 subtypes based on shared characteristics and evolutionary similarity. These are grouped... aeruginosa . Anti-CRISPR genes employ varied mechanisms to block CRISPR. Some prevent the CRISPR-Cas ...genome. Multiple Cas9 orthologs have also been characterized. Staphylococcus aureus (SaCas9) is smaller than... as guanosine, so the overall effect is an A->G change. Since this method targets RNA rather than DNA,...Xenopus References Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero DA, Horvath P....Wiedenheft B, Maxwell KL, Davidson AR. 2015. Multiple mechanisms for CRISPR-Cas inhibition by anti-CRISPR proteins...

...Neurodegenerative diseases are a group of disorders characterized by the progressive degeneration and ultimate... within 3-5 years from symptom onset. Disease mechanisms are still poorly understood but several genes... of nerve cells in the brain. Symptoms include changes in behavior and emotion, uncontrolled movements... -related antibodies including: Anti-KChIP1 K+ channel [K55/7R] Anti-Fig4/Sac3 [N202/7R] Anti-Ankyrin-...window) Rodent Models (Link opens in a new window) Michael J Fox Foundation (Link opens in a new window) A...therapies for those living with Parkinson's today. The Michael J. Fox Foundation has made a number of tools (Link...provided by the foundation can be found on our Michael J. Fox Foundation Plasmid Resource page . CHDI ...

...opsins, which are membrane channels and pores derived from algae and archaebacteria, that can be opened and...widely used Channelrhodopsin-2 (ChR2) from Chlamydomonas reinhardtii . ChR2 is a membrane channel that opens...cations to flow into the cell. Variants of channelrhodopsin have been engineered or identified to increase...microbial opsins, such as the Halorhodopsins, Archaerhodopsins and Leptosphaeria rhodopsins, are pumps that...

... 1 guanine nucleotide exchange factor VAV VAV2 vav 2 guanine nucleotide exchange factor - VAV3 vav 3 guanine... guanine nucleotide exchange factor FLJ40431 ZAP70 zeta-chain (TCR) associated protein kinase 70kDa FLJ17670...lab of Ian Frew . MXS Chaining Kit : From the lab of Pierre Neveu , the MXS-chaining system allows efficient...applications in mammalian systems. The MXS-chaining kit includes chaining vectors with multiple cloning sites...

...techniques for deletion, identification, and characterization. This strategy relies on cellular delivery... examine for scarring. This may be useful to characterize the non-deleted allele in monoallelic deletion...μl. This facilitates screening by allowing a multichannel pipette to be used for the remainder of the ...in each well) to a 96-well PCR plate using a multichannel pipette. Centrifuge PCR plate at 400 x g for...Validation of Biallelic Deletion Clones In order to characterize obtained clones and validate a successful knockout...and subject each clone to Sanger sequencing to characterize each deletion allele 5–7 . Repeating the PCR...Zhang F. Nature Protocols . 2013. PubMed . Characterization of genomic deletion efficiency mediated by...

... protein 35 ARHGEF2 Rho/Rac guanine nucleotide exchange factor 2 CCND CCND1 CCND2 CCND3 Cyclin D CDK CDK4...MLST8 MTOR associated protein, LST8 homolog MTOR Mechanistic target of rapamycin NF1 Neurofibromin 1 NFE2L2...Phosphatidylinositol-3,4,5-trisphosphate-dependent Rac exchange factor 2 PRKA PRKAA1 PRKAA2 PRKAB1 PRKAB2 PRKAG1... RAPGEF RAPGEF1 RAPGEF2 Rap guanine nucleotide exchange factor RAS KRAS MRAS NRAS Kirsten rat sarcoma ...protein SOS SOS1 SOS2 SOS Ras/Rho guanine nucleotide exchange factor SPRED SPRED1 SPRED2 SPRED3 Sprouty related...Tuberous sclerosis VAV1 Vav guanine nucleotide exchange factor 1 YAP1 Yes associated protein 1 Return ...

...activity, like transcription. Michael Davidson FP Collection Browse the Michael Davidson comprehensive collection...protein-protein interactions or conformational changes within a protein. Biosensors Monitor small biomolecules...protein of interest resides by using a well-characterized fluorescent fusion protein. In Vivo Imaging ... and UCSF Molecular Expressions Galleries from Michael Davidson and FSU Spectra Database hosted at the...

...ideally less than 10bp away. For small (<50bp) changes, you should consider using ssDNA oligo as donor...80bp of homology arms on each side flanking the change you'd like to introduce. We use Ultramer oligo ...the Cas9/sgRNAs vector (like PX330). For large changes (>100bp insertions or deletions), we typically ...to mutate the PAM 'NGG' in the HR template, like change it to 'NGT', or 'NGA', or 'NGC' (if it's within...required for successful targeting and cleavage, changing it to'NGT' or 'NGC', in addition to mutations ...typically use PAGE purified long oligos. For large changes (>100bp insertions or deletions), the Zhang lab...

...nlsdTomato Control Fishell 213829 pAAV_BiCHATe27_dTomato_nlsdTomato BiCHATe27 dTomato/nlsdTomato Control Fishell...BiSSTe10 Activator ChR2 Fishell 213830 pAAV_BiCHATe27_ChR2_mCherry BiCHATe27 Activator ChR2 Fishell 213856 pAAV_BiVIPe4...Link opens in a new window) and was adapted from Challis et al. (2022). Text version of the AAV decision...Viviana Gradinaru and cite as follows: PHP.eB, PHP.S: Chan et al., 2017 Pubmed 28671695 (Link opens in a new...