...219444 Anti-KChIP1 [K55/9R] KChIP1 Rat Mouse IgG2a 219445 Anti-KChIP1 K+ channel [K55/59R] KChIP1 K+ channel...190284 Anti-KChIP3 K+ channel [K66/27] KChIP3 K+ channel Rat Mouse IgG2a 190285 Anti-KChIP3 K+ channel ... 206760 KChIP3 K+ channel scFv [K66/27] K66/27 scFv KChIP3 K+ channel Rat Mouse 206761 KChIP3 K+ channel...38 scFv KChIP3 K+ channel Rat Mouse 206762 KChIP3 K+ channel scFv [K66/59] K66/59 scFv KChIP3 K+ channel...Human Mouse IgG2a 177442 Anti-Pan-KChIP K+ channel [K55/82R] Pan-KChIP K+ channel Mouse IgG2a 177443 Anti-CASK...CASK Human Mouse IgG2a 177444 Anti-KChIP2b K+ channel [K60/73R] KChIP2b K+ channel Rat Mouse IgG2a 177445...external) Human Mouse IgG2a 188175 Anti-KChIP2b K+ channel [K60/87R] KChIP2b K+ channel Rat Mouse IgG2a 188176...

...Locus-specific ChIP technologies consist of insertional chromatin immunoprecipitation (iChIP) and engineered...chromatin immunoprecipitation (enChIP; Fujita & Fujii, 2013). In iChIP, specific genomic regions tagged... Fujii Lab web page . Figure 1: iChIP general scheme Figure 2: enChIP general scheme Fujii Lab Plasmids...3xFLAG-dCas9/pCMV-7.1 enChIP plasmid, expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify...8xLexA-binding elements/pMD20 iChIP plasmid, LexA-binding elements 48874 3xFNLDD/pCMV-7.1 iChIP plasmid, expresses...3xFNLDD/pMXs-I2 iChIP plasmid, expresses 3xFNLDD in mammalian cells 48972 3xFNLDD/pMXs-IG iChIP plasmid, expresses...protein for enChIP analysis. Fujita T, Kitaura F, Oji A, Tanigawa N, Yuno M, Ikawa M, Taniuchi I, Fujii ...

...Elsaesser 140020 pAS_4xMma PylT_FLAG-AbKRS-chIPYE chimeric PylRS Chimeric photo-crosslinking-lysine Mammalian...translation machinery. To expand the genetic code, 4 major changes to the standard translation machinery are ...Bacterial Jason W Chin 174078 pDule-3-nitroTyrosine (A7) 3NY (A7) synthetase M. jannaschii 3-nitroTyrosine...Acd-A9 M. jannaschii acridone Bacterial TAG Ryan Mehl 197652 pDule2 - Mj Acd A9 Acd-A9 M. jannaschii acridone... sets. Many of these sets are derived from M. jannaschii , M. barkeri , or E.coli and can be mutated ...PI 31186 pEVOL-pAzF p-azidohenylalanine RS M. jannaschii p-azido-l-phenylalanine Bacterial TAG Peter Schultz...31190 pEVOL-pBpF p-benzoylphenylalanine RS M. jannaschii p-benzoyl-l-phenylalanine Bacterial TAG Peter...

...iCre CAG Mammalian Capecchi 113849 pCAGGS-pac-T2A-iCre iCre CAG Mammalian Capecchi 116879 CAG-Cremyc-2A-GFP...Mammalian Capecchi 125749 pCAG-sfGFP-iCre-ERT2 iCre-ERT2 - Tamoxifen inducible CAG Mammalian Capecchi 125821...Mammalian Capecchi 125822 pCAGGS-mTagBFP2-T2A-iCre (v2) TagBFP, iCre CAG Mammalian Capecchi 126006 pAAV2...Mammalian Cepko 69916 pAAV.cTNT.iCre iCre and tdtomato Chicken cardiac troponin T AAV Pu 70120 pEMS2159 iCre with...unskippable linker) - Tamoxifen inducible CAG Mammalian Capecchi 128167 pCAGGS-mTagBFP2-T2A-sfGFP-GSAx9-iCre-ERT2...GSAx9 linker) - Tamoxifen inducible CAG Mammalian Capecchi 128168 pCAGGS-mTagBFP2-T2A-iCre-ERT2 iCre-ERT2...-terminus) - Tamoxifen inducible CAG Mammalian Capecchi 128171 pCAGGS-mTagBFP2-T2A-iCreERT2 (PV) iCre-...

...Purify Genomic Regions The enChIP (engineered DNA-binding molecule-mediated ChIP) system, developed by the...the DNA, to create a DSB enChIP En gineered DNA-binding molecule-mediated ChIP , using a tagged-dCas9+gRNA...the ways the CRISPR components are leveraged to achieve specific goals, such as targeting multiple genomic...HDR) pathway. HDR relies on copying DNA from a matching template to accurately repair or fill in the missing...gRNAs into a single plasmid — though some have achieved targeting in the double digits. These multiplex...nickase system is utilized in many CRISPR systems to achieve specific genetic edits. The dCas9 molecule binds...or G, Y = C or T SpRYc - SpRY PAMs plus NNG; a chimeric version of SpRY Though these PAM-flexible Cas9s...

... Special considerations One type of ChIP assay, called native ChIP, does not use cross-linking and instead...accessible to antibodies. Detection of sdAbs is usually achieved by conjugation of a signaling molecule directly...can be detected either digitally by an imaging machine or by X-ray film. HRP cannot be used to distinguish...detect simultaneously. Conjugation The process of attaching signaling molecules to an antibody is done through...Immunoprecipitation (IP) Chromatin immunoprecipitation (ChIP) Visualization - used to visualize protein location...either X-ray film in a darkroom or an image capture machine. Special considerations The primary antibody must...immunoprecipitation (IP) and chromatin immunoprecipitation (ChIP) allow you to isolate specific proteins or protein...

...Gia Voeltz 89472 GFP-hChibby1 Mother Centrioles / Ciliary Base Chibby1 GFP Ken-Ichi Takemaru 89770 pLenti-EGFP-hChibby1...pLenti-EGFP-hChibby1 Mother Centrioles / Ciliary Base Chibby1 EGFP Ken-Ichi Takemaru 118084 pLenti-EB1-EGFP...EB1 EGFP Ken-Ichi Takemaru 122871 GFP-hCCDC11 Centriolar satellites CCDC11 EGFP Ken-Ichi Takemaru 54234... Cooper *Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein... Bement *Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein...Fanning *Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein... Pagano *Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein...

...Xenopus/C.elegans/Sea urchin pCS2+8NCerulean - Zebrafish/Xenopus/C.elegans/Sea urchin mCerulean N1 - Mammalian...Xenopus/C.elegans/Sea urchin pCS2+8NeGFP - Zebrafish/Xenopus/C.elegans/Sea urchin pKT0128 - Yeast Expression...Xenopus/C.elegans/Sea urchin pCS2+8NmCitrine - Zebrafish/Xenopus/C.elegans/Sea urchin pET mCitrine LIC cloning...Xenopus/C.elegans/Sea urchin pCS2+8NmOrange - Zebrafish/Xenopus/C.elegans/Sea urchin mOrange-N1 - Mammalian...Xenopus/C.elegans/Sea urchin pCS2+8NmCherry - Zebrafish/Xenopus/C.elegans/Sea urchin pET mCherry LIC cloning... mCherry, with neomycin cassette Zebrafish, Sea urchin, Xenopus, and C. elegans Hamdoun Lab Plasmids -...

...targeting chiRNAs Targeting chiRNAs are easily cloned by annealed oligos into the pU6-BbsI-chiRNA plasmid...low copy number. 45946 pU6-BbsI-chiRNA : Plasmid for expression of chiRNA under the control of the Drosophila...generated within a month. phsp70-Cas9 and a single chiRNA plasmid can be coinjected to generate mutations...Coinjection of phsp70-Cas9 and plasmids encoding two chiRNAs can induce defined deletions between the two cleavage...

...) , human CD2 (pSIR-hCD2) PDF 105.8 KB Fujii iChIP/enChIP to purify genomic DNA FLAG tagged dCas9 PDF ...Arabidopsis, C. elegans , sea squirt, cavefish, Chinese hamster, fruit fly, human, rice fish, mouse, silk...sequences. Organisms include human, mouse, rat, chicken, frog, zebrafish, fly, worm, Japanese rice fish...Currently supports: Human, mouse, rat, marmoset, pig, chicken, frog (X. tropicalis and X. laevis), zebrafish,...elegans, mouse, human, rat, rabbit, pig, possum, chicken, dog, mosquito, and stickleback. E-CRISP Identifies...design and cloning; injection protocol pU6-BbsI-chiRNA ; phsp70-Cas9 PDF 106.5 KB Orkin and Bauer Protocol...

... Angew Chem Int Ed Engl. 2018 Jun 27. Tetsuya Kitaguchi ATP Variants of ATP sensor iATPSnFR1.0 and iATPSnFR1.1.... PLoS One. 2014 Jun 24;9(6):e100252. Tetsuya Kitaguchi cAMP (cyclic AMP) Red fluorescent protein-based...imaging. Sci Rep. 2017 Aug 4;7(1):7351. Tetsuya Kitaguchi cAMP (cyclic AMP) Fluorescent sensor of cyclic...PDE5alpha. ACS Sens. 2017 Jan 27;2(1):46-51. Tetsuya Kitaguchi cGMP (cyclic GMP) FlincG3 (GFP-based cGMP sensor...PLoS One. 2014 Jun 5;9(6):e99095. Joseph Santos-Sacchi Chloride (Cl-) ClopHensorN chloride sensor (cytosolic...Anal Chem. 2019 Apr 2;91(7):4821-4830. Tetsuya Kitaguchi Glucose Red fluorescent glucose indicators for...ratio changes in different cellular compartments. Biochim Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519...

...particular gRNA. In the enChIP (engineered DNA-binding molecule-mediated ChIP) system, catalytically inactive... used for enChIP, as well as an anti-Cas9 antibody. Biotin tagging of dCas9 can be achieved by fusing ...researchers have expanded chromatin immunoprecipitation (ChIP) to allow purification of any genomic sequence specified...

...from our 'Chimeric RNA' design (meaning the mature processed form of crRNA is fused chimerically to a mature...experiments, consider that wildtype Cas9 with optimized chimeric gRNA has high efficiency but has been shown to...which has comparable efficiency to the optimized chimeric design but with better accuracy (in other words...nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will together...site with efficiency similar to using a single chimeric gRNA. At the same time, the off-target effects...

...MDH-related research into their teaching laboratories. They encourage instructors teaching different types of courses...projects, find teaching resources and protocols, learn about regional hubs, access a teaching mentor, and...development, membership to a local hub of educators teaching MDH CUREs, and opportunities to mentor future ...

... Description Category PI 157970 pTR-UF11 chimeric CMV/Chicken Beta actin (CBA) GFP Control Sergei Zolotukhin...subretinal injection. It was identified by John Chiorini and characterized by Shannon Boye’s lab. AAV44.9...Recombinant Adeno-Associated Virus Vector Expressing Retinoschisin. Hum Gene Ther Clin Dev . Sep;26(3):165-76. ...in future publications, please acknowledge John Chiorini and Shannon Boye and cite Boye, et al. 2020. Novel...

...nucleus accumbens medial shell and behavioral switching. Neurochemistry International 129:104482. PubMed...opens in a new window) Yu K, Ahrens S, Zhang X, Schiff H, Ramakrishnan C, Fenno L, Deisseroth K, Zhao ...in the Periaqueductal Gray Facilitate the Itch-Scratching Cycle via Article Tac1-Expressing Neurons in ...in the Periaqueductal Gray Facilitate the Itch-Scratching Cycle via Descending Regulation. Neuron 101(1...hypothalamus-habenula circuit controls aversion. Mol. Psychiatry 24(9):1351-1368. PubMed (Link opens in a new ...

... and its associated proteins or RNAs through ChIP (enChIP) or biotin-streptavidin pulldown (CAPTURE.) ...molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Biochem Biophys Res Commun . 439...for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product....Myers RM. 2015. CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins. Genome Res . 25(10)...

...zebrafish research. China Zebrafish Resource Center (CZRC) - A resource center based in China focused on collecting...Sean Megason Lab Zebrabow - Albert Pan and Alex Schier Labs. Multicolor fluorescent labeling in zebrafish...lineages and cell types in the vertebrate brain - Alex Schier Lab Other Tools of Interest A few examples of popular...

...S. pyogenes Virmilion Bullock and Port pU6-BbsI-chiRNA 45946 Drosophila BbsI none S. pyogenes O'Connor-Giles...Mammalian/Lentiviral BbsI none S. pyogenes Puro Ochiai pSB700 64046 Mammalian/Lentiviral none S. pyogenes...methylation S. pyogenes Puro Zoldos pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) 71707 Mammalian hU6 yes...Drosophila Utilizes endogenous tRNA processing machinery to excise multiple functional gRNAs from a single...simultaneously repressing multiple target genes in Escherichia coli. Koffas Do you have suggestions for other...

... system - Jaenisch Locus-specific ChIP technologies (iChIP & enChIP) using dCas9 - H. Fujii Disease Research...

... or 20 matching nucleotides, the second binds to mcherry with either 0, 11, 18, or 20 matching nucleotides...possible to study the effect of the amount and stoichiometry of enzymes. The pLC97 plasmid allows for quick...

...Parkinson's Lynda Chin 21691 pLenti6/V5/DEST-VPS35 VPS35 V5 CMV Parkinson's Lynda Chin 21827 TLS 1: hTLS.pBSKS...Gabriele Kaminski Schierle 108866 pT7-7 aSyn C141 SNCA T7 Parkinson's Gabriele Kaminski Schierle 108867 pET29b...Kaminski Schierle 108868 pLJM1 EGFP-Tau MAPT GFP CMV Parkinson's, FTD Gabriele Kaminski Schierle 109321 ... mScarlet CMV ALS Kyoko Chiba 184826 pSN838 KIF5A mScarlet CMV ALS Kyoko Chiba 184827 pSN526 KIF5A mScarlet...Strep polH ALS Kyoko Chiba 184832 pSN840 KIF5A mScarlet Strep polH ALS Kyoko Chiba 185714 pAAV-hSyn-GFP-IRES-mSNCA...pMXs-mPKR-FLAG-IG EIF2AK2 Flag LTR VWM disease Seiichi Oyadomari 101848 pTO_CHMP2B_LAP CHMP2B LAP CMV ...p426-Cup1p-FUS-FusionRed FUS FusionRed CUP1 ALS Michihiko Kataoka 188460 pKDR-Lenti-CBh.6-SOD1 SOD1 CBh ...

...Fab fragment libraries from chickens Methods for the generation of chicken monoclonal antibody fragments...antibodies as Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. Functional ...

...using combinations of AAV-based molecular tools to achieve targeted neuronal manipulation (aka intersectional...Potential combination with other technologies to achieve different experimental goals. How does this method... (that your group is presenting on) is used to achieve targeted neuronal manipulation. Figures, tables...

...-Puro 73039 Expresses Nluc and copGFP from the chicken beta-actin CMV enhancer pCDH-PGK-Nluc-P2A-copGFP-T2A-Puro... the EF1 promoter while pICPIS-CB contains the Chicken-beta actin promoter with CMV enhancer. pAdx-CMV-LacZ...polyA pICPIS-CB 73356 AdenoX shuttle vector with chicken-beta actin CMV enhancer promoter SV40 polyA...

...ddPCR are generally 2–3 fold higher than those achieved using the standard qPCR method. Titering by the...preparations is assayed by comparing the relative stoichiometric ratios of the viral capsid proteins VP1, VP2...Transfer Plasmid Addgene uses a rigorous barcode matching system that follows the sample from the DNA tube...

... in: A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation... in: Enhanced phosphoserine insertion during Escherichia coli protein synthesis via partial UAG codon ... described in: Expanding the genetic code of Escherichia coli with phosphoserine. Park HS, Hohn MJ, Umehara...

...and 63AA, respectfully). This next generation architecture has been shown to increase mutation induction...is driven by the strong CAG promoter or can be achieved by in vitro mRNA synthesis from the T7 promoter...cells, (iv) an improved, truncated TALE backbone architecture as established by Miller et al. (PMID: 21179091...

...sensitivity, wide dynamic range, and resistance to photobleaching has made luciferase a common choice in assays... of renilla luciferase Reuben Shaw 48688 pUltra-Chili-Luc Firefly Ubc Lentiviral expression of firefly... Mammalian expression of firefly luciferase Ming-Chih Lai 115366 p CIneo-RL Renilla CMV Mammalian expression...

...Resources iGEM - International Genetically Engineered Machine Foundation Registry of Standard Biological Parts...synthetic biologists SEVA - Standard European Vector Architecture Open Plant - Plant SynBio resource EraSynBio...

...none 2, 5 Richie 121539 pAAV SYN1 HA-hM3D(Gq) hM3D(Gq) - Activation HA fusion none 2, 5 Richie 50467 pAAV-CaMKIIa-HA-hM4D...

...CRISPRi), works independently of host cellular machineries, requiring only a nuclease-deficient Cas9 (dCas9...binding specificity is determined jointly by 20-bp matching region on the sgRNA and a short DNA motif (protospacer...

... and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate...molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Biochem Biophys Res Commun. 2013...

...scientific review of our viral pages, we thank Boston Children's Hospital Viral Core . Viral Plasmid Biosafety...Au S, Panté N. How viruses access the nucleus. Biochim Biophys Acta. 2011 Sep;1813(9):1634-45. doi: 10.1016...

... and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate...using streptavidin affinity purification. For both enCHIP and CAPTURE, design your gRNA sequence to direct...

...strategy relies on cellular delivery of a pair of chimeric single guide RNAs (sgRNAs) to create two double...of PCR products. Finney M, Nisson PE, Rashtchian A. Current Protocols in Molecular Biology. 2001...

...protein reporter genes in Gram-negative bacteria. Hochschild Bacterial Two-Hybrid System Various Plasmids ...with lambda-C1 operator Beta-galactosidase Ann Hochschild Bacterial two-hybrid assay using pBRα-β flap ...

... Acad Sci U S A. 2014 Jul 22;111(29):10678-83. Capecchi Replicating EBNA1 episome Human Fluorescent-tagged...reprogramming factors Fluorescent tagged episomals for stoichiometric induced pluripotent stem cell reprogramming...

...Constitutive 9 Patriarchi 187180 pAAV_hSyn1_nLightR Syn nLightR none Constitutive 9 Patriarchi Oxytocin (OT...

...assemble TAL effector repeat arrays on a single architectural framework and the DNAs produced by all three...