... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from...were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell...) alone at 1.1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct...

...resistant cell types. CreLite - George Eisenhoffer Lab. An optogenetically controlled Cre/loxP system that ...toolbox of genetic methods, such as Tol2 transposons, Cre-mediated site-specific recombination, morpholinos...fluorescent proteins, reporters, biosensors, and Cre expression plasmids. Large collections or kits are...that are amenable to genetics, high-throughput screening and non-invasive live imaging. A standard toolbox... Amro Hamdoun Lab. A useful “starter kit” for screening fluorescent protein fusion expression in organisms...antibodies to zebrafish cell surface receptors and secreted proteins. Return to top Other Resources Addgene...American Zebrafish Network (LAZEN) - LAZEN aims to increase the use of zebrafish as a model organism and to...

... pSico 3rd Conditional (Cre-lox), stable expression of shRNAs; addition of Cre turns on shRNA expression...expression. See article for more cre-lox shRNA expression variants. Jacks 19319 pLJM1-EGFP 3rd CMV-driven EGFP...systems available at Addgene have been designed for increased researcher safety. Read our Lentiviral Guide for...22502 pCEP4-tat N/A Packaging Expression of tat; increases viral titer Kasparov 22501 pHEF-VSVG N/A Envelope...

...9:00 AM Breakfast and Welcome Melina Fan 9:30 AM Cre and Flpo Katherine Matho 10:15 AM AAV Retrograde ...and Other Forms of Regulation Ben Deverman 2:00 PM Cre-DOG and Combinatorial Strategies Connie Cepko 2:45...

...Channelrhodopsins, Halorhodopsins, Archaerhodopsins Recombinases Cre, Flp, Dre Biosensors Calcium Sensors, Glutamate Sensors...

...Genome Engineering Genome Engineering Overview CRISPR Cre-lox TALENs Zinc Fingers Luminescence Fluorescent ...

...inhibition. For example, search for "ChR mCherry", "Cre inhibition", "Boyden 2014", or other terms. Looking...channelrhodopsin have been engineered or identified to increase the spectral and kinetic properties as well as...

...EGFP Del Bene pUAS:Cas9T2ACre;U6:sgRNA1;U6:sgRNA2 74010 Zebrafish U6 yes, cut S. pyogenes Cre Del Bene pTE4396...pyogenes mCherry Kuhn AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR 60229 Mammalian/AAV SapI none S. pyogenes... pyogenes Zhang AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR 60231 Mammalian/AAV ...pyogenes Zhang AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR 60226 Mammalian/AAV SapI none S. pyogenes...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...activity, experimental considerations, and more, created and maintained by the Doudna Lab . Plasmid Expression...

...RNAi, empty backbones, fluorescent proteins, and Cre expression plasmids. Large collections or kits are...

...Noteworthy: AAV.rTH.PI.Cre.SV40 ready-to-use viral prep useful for dopamine research. CBA-driven, Cre-dependent...blood brain barrier integrity, and using CRISPR screens to understand more globally how neurons function...antibodies and scFvs for neuroscience research created with high-volume hybridoma sequencing on the NeuroMabSeq...community. This collection highlights plasmids created with support from the BRAIN Initiative. Jackson...Link opens in a new window) . This resource was created with support from the Chan Zuckerberg Initiative...

...vector assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general cloning plasmid..._CEBPA_1 PX458_CEBPA_2 CREB1 Human FLAG pFETCh_CREB1 PX458_CREB_1 PX458_CREB_2 TGIF2 Human FLAG pFETCh_TGIF2...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...genome modification. This system can be used to create C- and N-terminal epitope tags. The plasmids in...has developed a systematic and scalable method to create marker-free mutations, insertions, and deletions...stem cell (hiPSC) lines. These cell lines were created using CRISPR and the donor plasmids containing ...

...Hynes Lab Plasmids - A system for Cre-regulated RNAi in vivo Dupuy Lab and... from the SP6 promoter pCS2 mt-GFP - Create myc-tag, GFP-tag fusions for transcription...some experiments, scientists often want to create stable cell lines in which the plasmid is ...

...pCFJ90 (pharyngeal co-injection marker); pDD104 (Cre recombinase) PDF 102.5 KB Jaenisch CRISPR plasmid.... Green Listed - a CRISPR Screen Tool Design gRNAs for custom CRISPR screens targeting a long or short...labs who have deposited CRISPR plasmids , we've created a series of links and guides to help you use CRISPR...considerations, and more. CasPEDIA is a community resource created and maintained by the Doudna Lab at the Innovative...elegans, mouse, human, rat, cow, dog, pig, Thale cress, rice (Oryza sativa), tomato, corn, monkey (macaca...additional HDR & gRNA plasmids are available for CREB1, ATF1, GABPA, & RAD21 PDF 134.7 KB Musunuru CRISPRs...

...PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Renilla EF1α AAV9-CRISPR knockout ...promoter/enhancer regions. Blue/white screening can be used for screening of transformants in cloning process...transcriptional reporters. However, they are not secreted, so cells must be lysed or the substrate must ...to measure luminescence. Gaussia luciferase is secreted and more stable, making time course experiments...size. NanoLuc® Luciferase is small, bright, and a secreted version of the protein is available – these attributes...luciferase mutants : A set of three sensors that are increasingly structurally destabilized versions of Firefly.../green light and red light, respectively, for increased usefulness in in vivo applications. LuxSit-i :...

...Lenti-sgAtm#1/Cre ATM CMV Ataxia telangiectasia Charles Swanton 173580 Lenti-sgAtm#2/Cre ATM CMV Ataxia...Lewy body dementia Tiago Outeiro 89643 Lenti-sgAtm/Cre ATM U6 Ataxia telangiectasia Monte Winslow 89755 ...T5 Parkinson's Suzanne Pfeffer 208392 Lenti-sgRNA-Cre-GpNLuc-sgmATM ATM Ataxia telangiectasia Gaorav Gupta...Spinocerebellar ataxia 19 James Trimmer 129409 Slc1a3-CreERT2 Targeting Vector SLC1A3 PGK Episodic ataxia Walker...

...used for large scale edits. These proteins, like Cre recombinase or phage derived serine integrases, insert... GAT; increase nuclease fidelity SpCas9-NG - NG; increase in vitro activity SpG - NGN; increase nuclease...others work to increase Cas9’s proofreading capabilities. No matter the method, increased fidelity enzymes...backbone HypaCas9 - increase Cas9 proofreading and discrimination evoCas9 - decrease off-target effects...domains to expand PAM recognition, increase specificity, and decrease off-target effects Sniper-Cas9 - ...compatible with truncated gRNAs to increase specificity SuperFi-Cas9 - increases fidelity with reduced nuclease...integration of donor DNA. Genome-Wide Screens Using CRISPR In a genetic screen, a population of cells with mutations...

...mutant allele of a gene ( point mutant )? Increase or decrease expression of a target gene? Once you have...High-fidelity Cas enzymes increase specificity. Dual-nickase approach increases specificity but is less ...efficiency. In fact, cleavage efficiency may increase or decrease depending upon the specific nucleotides ...alone for mammalian cell lines. Activate (CRISPRa) Increase expression of an endogenous gene(s) without permanently...vivo A common choice for conducting genome-wide screens using CRISPR AAV transduction CRISPR elements are...Transient expression of CRISPR components Expression decreases as RNA is degraded within the cell Short window...window of CRISPR component expression may decrease off-target effects Can be used to generate transgenic ...

...Upon activation, NK cells secrete perforin and granzyme B, two enzymes that create pores within an infected...LSIGN, MGC129964, MGC47866 CREB1 cAMP responsive element binding protein 1 CREB, MGC9284 CTSB cathepsin ...chromogranin A (parathyroid secretory protein 1) CGA CHGB chromogranin B (secretogranin 1) SCG1 CKLF chemokine-like... SDS, SWDS SCG2 secretogranin II (chromogranin C) CHGC, SN, SgII SCGB3A1 secretoglobin, family 3A, member...HIN1, LU105, MGC87867, PnSP-2, UGRP2 SCT secretin - SCTR secretin receptor SR SCYE1 small inducible cytokine... B cells mediate humoral immunity through the secretion of antibodies, and also function as antigen presenting...

... Bauer's labs details the use of CRISPR/Cas9 to create genomic deletions in mammalian cell lines. Along... pair of chimeric single guide RNAs (sgRNAs) to create two double strand breaks (DSBs) at a locus in order...strategies for genes and non-coding elements. For creating a gene knockout, two sgRNA located within exons...be 24-mer oligos ( Table 4 ). Design Deletion Screening Primers Design one set of primers internal to ... for future plating. Plate the other half for screening and primer validation (see step 6 ). NOTE: This... allows for sufficient cell proliferation for screening genomic DNA (gDNA) for the intended deletion by...visible with ~2 mm diameter. Primer Validation and Screening for CRISPR/Cas9-Mediated Deletion Isolate gDNA...

...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...double stranded breaks. Cytosine base editors are created by fusing Cas9 nickase or catalytically inactive...converted to thymidine through base excision repair, creating a C to T change (or a G to A on the opposite strand..., which is treated like guanosine by the cell, creating an A to G (or T to C) change. Adenine DNA deaminases...deaminases do not exist in nature, but have been created by directed evolution of the Escherichia coli TadA...evolved TadA domain is fused to a Cas9 protein to create the adenine base editor. Both types of base editors...adjust the editing window, or adding fusions that increase product purity such as the DNA glycosylase inhibitor...