...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...

...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 ...

...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...

... 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...

...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...

...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 ...