...preps SARS-CoV-2 protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins ...control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain...for targeted expression. Plasmid 75470 is now available in AAV1 for Flp-dependent expression of ChR2(H134R...bound to an mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression of ChR2 ...Lentiviral preps for expressing SARS-CoV-2 proteins ChR2 for targeted expression DREADDs GFP dependent...2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV...pAAV-Ef1a-fDIO mCherry Karl Deisseroth 112677 AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP Brandon...

...transgene expression across different cell types (Aschauer et al., 2013). Expression of hM4D(Gi)-mCherry from...single-vector systems typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be...stable Cas9-expressing cell line and then transduce this line with lentiviruses expressing various gRNAs...single-vector system). Tips for using this plasmid Cas9 expression is typically cytotoxic, which makes generation...-hM4D(Gi)-mCherry Our current AAV inventory consists almost entirely of viruses expressing DREADDs from... injected, the DREADD will only be expressed in cells that express cre recombinase. Typically, this feature...to attain cell-specific expression in transgenic animals, where cre expression is restricted to a particular...

...pAAV-hSyn-DIO-hM4D(Gi)-mCherry Controls Plasmid Serotype Name 50459 AAV1 pAAV-hSyn-DIO-mCherry 27056 AAVrg...pAAV-CaMKIIa-hChR2(H134R)-mCherry 26976 AAV5 pAAV-hSyn-hChR2(H134R)-mCherry 26973 AAVrg pAAV-hSyn-hChR2...them! DREADDs Plasmid Serotype Name  44361  AAV1 pAAV-hSyn-DIO-hM3D(Gq)-mCherry    44362  AAV1 ... AAV5 pAAV-Ef1a-fDIO mCherry 112677 AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP 50457 AAV2 pAAV-hSyn-DIO-EGFP...constructs from Christopher Harvey’s lab. These plasmids localize opsins to the soma to avoid off target...neighboring neurons. We have also packaged hChR2(H134R) plasmids from Karl Deisseroth’s lab, which includes pAAV-hSyn...H134R)-EYFP that is both Cre and Flp dependent. Plasmid Serotype Name 124603 AAV9 pAAV-EF1a-DIO-ChrimsonR-mRuby2...

...control RFP expression plasmid.  With this set up, all successfully transfected cells will express RFP while...using the supplied donor plasmid will express GFP. If there’s GFP expression, presumably the endogenous...locus using GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs..., can result in an in-frame shift of mCherry and thus expression. As a proof of concept the Denham lab...stem cells.  Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems...desired genomic edit.  Find SRIRACCHA plasmids at Addgene GEMCherry2, an improved monomeric fluorescent ...iteration was named GEmCherry1, due to the “genomic fragment inserted into mCherry”. However this one ...

...pAAV-FLEX-tdTomato pAAV-GFAP104-mCherry pAAV-mDlx-NLS-mRuby2 CAG-NLS-GFP pAAV-hSyn-mCherry Read on to learn more...pAAV-FLEX-tdTomato, pAAV-GFAP104-mCherry, pAAV-mDlx-NLS-mRuby2, CAG-NLS-GFP,and pAAV-hSyn-mCherry at Addgene. Viral ... about. Controls Plasmid Serotype Name 114472  AAV5, AAV8  pAAV-hSyn-mCherry 50459  AAVrg  pAAV-hSyn-DIO-mCherry...indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST Chemogenetics Plasmid Serotype...  pAAV-EF1a-mCherry-IRES-Flpo 55637  AAV1  pAAV-EF1a-Flpo Calcium sensors Plasmid Serotype Name...protein and control AAV These fluorescent protein expression vectors can be used to visualize cells and to...types. For example, the mDlx enhancer restricts expression to GABAergic interneurons. These vectors can ...

... comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry. ACE was used to...wild-type mCherry and a T2A site and expressed in cells. mCherry is constitutively expressed to identify... mutated mCherry and a downstream constitutively active eGFP. To create the inactive mCherry, the lab ...DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using Fluorescence-activated...dividing the number of eGFP and mCherry positive cells by the number of only mCherry positive cells (Martin et...efficiency of plasmid delivery to a cell. In contrast, TREE provides a read out for plasmid delivery and... (5’-TCA-to-TUA) found within the 43 base-pair mCherry insertion. Editing of these motifs generates lesions...

...promoter, and a mCherry reporter downstream of the PBAD promoter. To use this plasmid with your gene of...interest, you would replace mCherry with your gene of interest. pBLADEONLY: This plasmid contains only BLADE ...pTRC99a plasmid. It does not include the PBAD promoter. If you already have a pBAD33-based plasmid expressing...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They...-inducible AraC dimers in E. coli) that turns expression on and off with blue light.   Inspired by @UriAlonWeizmann...AraC and other promoter systems, head over to our Plasmids 101: Inducible Promoters article.) Modifying AraC...VVD and the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none...

...They expressed this OptoNB-SOS fusion protein in cells that expressed membrane-localized mCherry. With...these experiments.  AsLOV2 Inertion Site mCherry #1 mCherry #2 GFP Loop 1 dark-induced dark-induced... Resources on Addgene.org Find nanobody expression plasmids here Learn more about optogenetics in Addgene's...Toettcher Lab monitored how AsLOV2 insertion into mCherry or GFP nanobodies affected binding of their target... its co-localization with a membrane-localized mCherry or GFP protein. With this system, if the OptoNB...sites in all eight surface-exposed loops of an mCherry nanobody. Only insertion into loops 1 and 6 resulted...scheme to generate two more OptoNBs, one against mCherry and one against GFP. Insertion of AsLOV2 into loop...

...strong, ubiquitous expression. This is a Cre-dependent switch that expresses nuclear mCherry in the absence...is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available as 20...ubiquitous expression. pAAV-mDlx-NLS-mRuby2 (99130-AAVrg): The Dlx element directs expression in forebrain...pAAV-hSyn-hChR2(H134R)-mCherry (26976-AAV5) pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5) ...entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors and ... AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This collection...The Synapsin promoter directs broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG...

...FLEx switch that turns BFP expression off, while turning on mCherry expression. For this FLEx switch to ...of Cre recombinase. This plasmid now specifically drives expression of mCherry instead of BFP. Beyond switching...elements to turn off expression of one gene, while simultaneously turning on the expression of another! What...2003): Basic FLEx switch will express BFP in the absence of, or mCherry in the presence of Cre recombinase...allowing for expression of GeneR. This allows a researcher to test whether or not expressing GeneR alone...potential to express the transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic...ensures Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...

...114472  AAV5, AAV8 pAAV-hSyn-mCherry 50459  AAV9, AAVrg pAAV-hSyn-DIO-mCherry 37825 AAV1, 2, 5, 8, 9 ...AAVs New DREADDs Plasmid Serotype Name 75033  AAV1 pAAV CD68-hM4D(Gi)-mCherry 50467  AAV2 pAAV-...populations. We now offer a Flp-expressing AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase...pAAV-hSyn-EGFP Recombinases Plasmid Serotype Name 55634  AAV1  pAAV-EF1a-mCherry-IRES-Flpo 55637  AAV1...Chemogenetics Plasmid Serotype Name 119472  AAV5  AV SYN PSAM4 GlyR IRES EGFP   Biosensors Plasmid Serotype...detection PSAM4 GlyR for chemogenetic inhibition Flp expression for intersectional studies And more! Control...inventory of serotype testing AAV New control AAVs Plasmid Serotype Name 105530  AAV1 pAAV.CMV.PI.EGFP.WPRE.bGH...

...4x-FLAG-mCherry-H2B), cytoskeletal (middle, 4xFLAG-mCherry-β-actin), and mitochondrial (mito-mCherry-1xFLAG...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure...Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...

...promoter to drive expression of Cre-mCherry and a second promoter to drive expression of a Cre-dependent... reporter. Expression of GFP will only result in cells where Cre-mCherry is also expressed. If Cre-mCherry...to test for cell type-specific expression of GFP. Half (113) expressed in retinal cells four weeks post-injection...More than 90% of expression occurred in one type of cell More than 50% of expression occurred in one type...of targeted expression. Surprisingly, <1% of synthetic promoters replicated the expression specificity...restrict expression to a single cell type based on the overlap in cell types both promoters expressed in. ...Cre-mCherry expresses in photoreceptors and horizontal cells, and Cre-dependent-GFP expresses in horizontal...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...about a recent plasmid deposit please sign up here.   Listen this episode of Hot Plasmids! Light controls...available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius...reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry fluorescence is activated...Hahn's lab at the University of North Carolina (plasmids at Addgene). It consists of two domains, LOV2 ... the same number of fluorophores. You can find plasmids containing the cobalamin binding riboswitches ...

....org Find Cre-Lox Plasmids Find Optogenetics Plasmids Find Chemogenetics Plasmids ...networks Double inverted vectors expressing light inducible ion channels (plasmid # 20298 for example) have ... 1: Plasmid mix to label neuronal morphology (eYFP) and the synaptic protein PSD95 (PSD95-mcherry) under...all labeling plasmids and the low concentration of recombinase restricts their expression to individual... Figure 2: Expression of a morphological marker (eYFP) and synaptic marker (PSD95-mcherry), under the...specific) control of DNA recombination and gene expression can be advanced by the coordinated use of fellow...means to spatiotemporally control FLP and Cre expression have been developed. Read on to learn more about...

...transformed with the sgRNA cassette on a plasmid expressing mCherry and resistance selection marker HygR....of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...recombination methods). By using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus...selection. Using an inducible Cas9 expression and removing the sgRNA plasmid after identifying successful knockouts... of the target gene. Lastly, curing the mCherry-sgRNA plasmid leaves an edited strain carrying Cas9 to... or want to swap out the constant domains, the plasmids for the heavy chain and light chain are also available...difficult to study in vitro, requiring either expressing it with additional cofactors or using significantly...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent...cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s. b) Quantification... CRISPR plasmids are always being added to the repository. To find all of the CRISPR plasmids available... Xiao Wang Lab has recently deposited a set of plasmids that helps identify base edited cell populations...populations without end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter...

...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral...in the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from.... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...vectors are tested in vitro and in vivo for gene expression and/or function. These data are sometimes reported...Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at 1.7E6 viral genomes...mL, or both. Two days later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was...

...ubiquitous promoter drives expression of an mCherry reporter.  After using mCherry for FACS sorting of the...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ...region-specific, inducible expression.   Find heat-inducible lentiviral plasmids here!   Wu Y, et. al. Control... missed something smokin'? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ...and Cas13 were compared head-to-head for gene expression changes of non-target genes and RNA-seq of a ...Figure 3 Colognori et al 2022.   Find CRISPR Csm plasmids here! Colognori D, Trinidad M, Doudna JA. Precise...heat-inducible Hsp (heatshock promoter) drives expression of the tumor-seeking CAR (chimeric antigen receptor...

...Addgene #140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...transcription of the mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS...too high expression of these markers can lead to non-specific labeling. They generated plasmids tagged ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...