...native expression levels. However, it is advisable to first express the fusion protein from a plasmid-based...instance, Lifeact-mTurquoise2 (Plasmid #36201) or mScarlet-tubulin (Plasmid #85045). We often use restriction...mTurquoise2, mEGFP, mNeonGreen, mScarlet(-I), and mCherry. Keep the linker sequence short In the early days...strategy (Sheridan, 2002). Avoiding protein over-expression artifacts The introduction of DNA that encodes...of protein within a cell and may lead to over-expression artifacts. This is an important downside of the...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ...

...fused to a reporter gene (e.g., mCherry) to easily detect opsin-expressing cells. The resulting fusion gene... about FLEx vectors Download our Plasmids 101 eBook Read our plasmids blog posts Resources on the Addgene...molecular biology reference Find Cre-lox plasmids at Addgene Browse plasmids at Addgene ...not naturally light-sensitive, so the selective expression of opsin genes in targeted cells makes it possible...How to get an opsin into the brain In order to express opsins within the brain, researchers inject genetically...relatively stable extra-chromosomally long-term expression and low ability to induce innate immune responses... promoters; however, those usually drive weak expression of downstream genes. Strong cell-specific promoters...

...pAAV-hSyn-mCherry Syn mCherry Control Deisseroth 114469 pAAV-CaMKIIa-mCherry CamKII mCherry Control Deisseroth...pAAV-Ef1a-mCherry EF1a mCherry Control Deisseroth 114471 pAAV-Ef1a-fDIO mCherry EF1a mCherry, Flp-dependent...pAAV EF1a Nuc-flox(mCherry)-EGFP EF1a Color-flipping switch. Expresses NLS-mCherry in the absence of Cre...55634 pAAV-EF1a-mCherry-IRES-Flpo EF1a Flpo and physically-separate mCherry expression Recombinases Deisseroth...Deisseroth 50459 pAAV-hSyn-DIO-mCherry Syn mCherry, Cre-dependent Control Roth 59462 pAAV-CAG-tdTomato...24593 AAV-pgk-Cre PGK Cre expression Recombinases Aebischer 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a Cre and ...and physically-separate mCherry expression Recombinases Deisseroth 105540 pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40...

...Fluorescent Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...tag with a serious drawback. The standard EGFP plasmid that used to be sold by Clontech and is in a freezer...solution is to restrict imaging analysis to cells expressing the lowest levels of the fusion protein. This... which cannot form heterodimers (i.e. EGFP and mCherry). These simple strategies can help you make better...

...Collections INTRSECT Plasmid Collection INTRSECT Plasmid Collection INTRSECT Implementation Plasmids References ...Find intersectional expression plasmids that combine multiplexed recombinases, including Cre, Flp, and...Example of viral targeting of cells expressing Cre AND Flp, here expressed through a double transgenic animal...recombinase-expressing mouse lines 2–9 and combinations of transgenic recombinase-expressing animal lines...novel INTSRECT plasmids for groups interested in producing their own INTRSECT plasmids. Online spreadsheet... pAAV-Ef1a-mCherry-IRES-Cre None Yes 55637 pAAV-EF1a-Flpo None Yes 55634 pAAV-EF1a-mCherry-IRES-Flpo None...137132 pAAV-Ef1a-Con/Fon-mCherry Cre AND Flp Yes 137133 pAAV-Ef1a-Con/Foff 2.0-mCherry Cre AND NOT Flp FRT/...

...SepOTSγ Plasmid 68285 SepOTSδ Plasmid 68286 SepOTSϵ Plasmid 68287 SepOTSζ Plasmid 68288 SepOTSη Plasmid...SepOTSθ Plasmid 68290 SepOTSι Plasmid 68291 SepOTSκ Plasmid 52054 SepOTSµ ( aka. B40 OTS) Plasmid 68294 ...68294 SepOTSν Plasmid 68295 GFP E17TAG Plasmid 68296 GFP S2TAG Plasmid 68297 GFP Q157TAG Plasmid 68298 GFP...Amp) Plasmid 68300 MBP-MEK1 Plasmid 68301 MBP-MEK1 S218TAG Plasmid 68302 MBP-MEK1 S222TAG Plasmid 68305...pNAS1b split mCherry 14-3-3β ctrl Plasmid 112030 pNAS1b split mCherry NEDD4 WW2 neg ctrl Plasmid 112029 pNAS1b...pNAS1b split mCherry NEDD4 WW2 pos ctrl Plasmid 111883 pNAS1b split mCherry NEDD4-2 WW2 Plasmid 111882 pNAS1b...pNAS1b split mCherry NEDD4 WW2 Plasmid 111881 pNAS1b split mCherry 14-3-3σ Plasmid 111880 pNAS1b split...

... Plasmid Collections Cre-lox Plasmids Cre-lox Plasmids Additional...preparations of plasmids that are currently in our repository. ID Plasmid Description Promoter Expression System...Hirrlinger 107312 AAV-hSyn-mCherry-P2A-Cre-WPRE mCherry and Cre; expressed in neurons hSyn AAV Yang 107313...107313 AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA mCherry and Cre; expressed in excitatory neurons aCamKII...interest: ID Plasmid Description Expression System PI 11578 pSico Cre activates shRNA expression, removal ... 26888 pmCherry-CRY2-CreN CRY2-CreN and mCherry coexpression; Light inducible; Use with plasmid 26889.... 26889 pmCherry-CIBN-CreC CIBN-CreC and mCherry coexpression; Light inducible; Use with plasmid 26888....

...Gradinaru 213945-PHPeB pAAV_BiSSTe4_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...Gradinaru 213941-PHPeB pAAV_BiPVe3_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...Gradinaru 213937-PHPeB pAAV_BiPVe4_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...Gradinaru 213915-PHPeB pAAV_BiLAMP5e3_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...Gradinaru 213856-PHPeB pAAV_BiVIPe4_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...Gradinaru 213830-PHPeB pAAV_BiCHATe27_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...Gradinaru 213815-PHPeB pAAV_BiSSTe10_ChR2_mCherry AAV construct expressing mCherry-tagged channelrhodopsin...

... on the plasmid. gRNA plasmids that do not co-express a Cas protein require a separate plasmid that does...Vectors CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut...maintained by the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection...CBh-Cas9-T2A-mcherry-P2A-Ad4E1B 64221 Mammalian see plasmid page yes, cut S. pyogenes mCherry Kuhn pU6-(...CBh-Cas9-T2A-mcherry-P2A-Ad4E4orf6 64222 Mammalian see plasmid page yes, cut S. pyogenes mCherry Kuhn AAV:...can be used to express multiple gRNAs from a single plasmid with or without expression of Cas9. Highlighted.... Can express Cas9 with up to 3 gRNAs. Frew pSQT1313 Mammalian Single plasmid for the expression of multipe...

...driven KOR DREADD expression. Bryan Roth 75033-AAV1 pAAV CD68-hM4D(Gi)-mCherry Expression of hM4D(Gi) using... 83898-AAV1 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic interneurons... 83898-AAV9 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic interneurons...83898-AAVrg pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic interneurons...Chemogenetics expression plasmid Scott Sternson 123308-AAV9 pAAV-hSyn-GRAB_NE1m Expresses the genetically-encoded...135634-AAV1 pAAV-S5E2-ChR2-mCherry AAV vector to drive the expression of ChR2-mCherry in PV cortical interneuronsunder...135634-AAV9 pAAV-S5E2-ChR2-mCherry AAV vector to drive the expression of ChR2-mCherry in PV cortical interneuronsunder...

...AAV-EF1a-BbChT EF1a mCherry and mTFP Cre dependent 9 Sanes 114471 pAAV-Ef1a-fDIO mCherry EF1a mCherry Flp dependent...Constitutive 5 Zeng 58909 pAAV-GFAP104-mCherry GFAP104 mCherry Constitutive 5 Boyden 59462 pAAV-CAG-tdTomato...Constitutive 2 Patrick 114469 pAAV-CaMKIIa-mCherry CaMKIIa mCherry Constitutive 1, 2, 5, 8, 9, rg*, PHP.eB...PHP.eB Deisseroth 114470 pAAV-Ef1a-mCherry EF1a mCherry Constitutive 1, 2, 5, 8, 9, rg*, PHP.eB Deisseroth...Deisseroth 114472 pAAV-hSyn-mCherry hSyn mCherry Constitutive 1, 2, 5, 8, 9, rg* Deisseroth 117382 hSyn1-eYFP ...112677 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP EF1a NLS-mCherry or nls-EGFP Cre dependent 1, 2, 5, ...1, 2, 5, 8, rg* Roth 50459 pAAV-hSyn-DIO-mCherry hSyn mCherry Cre dependent 1, 2, 5, 8, 9, rg* Roth 51502...

...6B) - Mammalian Expression pLV-mCherry - Mammalian Lentiviral Expression pCS2+8CmCherry - Zebrafish/Xenopus...pBad-EBFP2 - Bacterial Expression EBFP2-N1 - Mammalian Expression EBFP2-C1 - Mammalian Expression EBFP2-pBAD - ...- Mammalian Expression pCyPet-C1 - Mammalian Expression pCyPet-His - Bacterial Expression CYPet-N1 - Mammalian...Bacterial Expression iLOV-N1 - Mammalian Expression iLOV-C1 - Mammalian Expression AcGFP1 475 505 28 Prone ... - Mammalian Expression sfGFP-C1 - Mammalian Expression sfGFP-pBAD - Bacterial Expression moxGFP 486 510...Mammalian Expression pCAG-GFP - Mammalian Expression pLV-eGFP - Mammalian Lentiviral Expression Ac5-STABLE1...1 - Mammalian Expression mEGFP-N1 - Mammalian Expression mEGFP-C1 - Mammalian Expression mEGFP-pBAD - ...

...CRISPR Guide Find Plasmids for Cas9 Purification Find CRISPR Plasmids for Mammalian Expression ...editing. How to use Addgene plasmids for Easi-CRISPR You can use Addgene plasmids to create a repair template...of Addgene plasmid options for templates - see the example at right using blue flame plasmid pmScarlet_...desired insert (e.g., mScarlet).   Plasmid ID Plasmid Name Tags 69090 pMJ915 MBP (N ...pre-CRISPR was very arduous. First, you’d construct a plasmid with your desired changes, long homology arms and...template! If you have a plasmid with your desired insertion feature (e.g., a fluorescent protein), you can...cassettes, such as fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre, Flp etc.), and tetracycline...

...From the Bikard Lab, CRISPR plasmid collection for modulating expression levels of two targets independently... can obtain a range of expression levels. This collection comprises 20 plasmids, each carrying two CRISPR...binds to mcherry with either 0, 11, 18, or 20 matching nucleotides. Together, these plasmids allow to ...range of expression levels for these two reporters simultaneously. By fusing sfGFP and/or mCherry to genes... modulate expression of two genes without cross‐talk. (A) Schematic of the strain expressing two reporters...the two reporters, mCherry and sfgfp in this case, can be controlled using a plasmid‐borne CRISPR array...Repression Collection Bikard Lab CRISPR Repression Plasmids You may also like... CRISPR Guide CRISPR Protocols...

...which can be used to control gene expression. See our Cre-lox Plasmids page for more information about ..., 2, 5, 8, 9 Wilson 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a mCherry (not a fusion tag) 8, rg* Deisseroth... Syn none 9 Wilson 107312 AAV-hSyn-mCherry-P2A-Cre-WPRE Syn mCherry 1 Yang 107738 pAAV-hSyn-Cre-P2A-dTomato...none rg* Wickersham 55634 pAAV-EF1a-mCherry-IRES-Flpo EF1a mCherry (not a fusion tag) 1, rg* Deisseroth...We provide high quality AAV preps from select plasmids in the repository. Browse our AAV collection of...retrograde serotype, produced with the AAV retro helper plasmid from the Karpova and Schaffer labs. See the Retrograde...options. Select from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need....

...below for CRISPR prime editing plasmids. Plasmids are available for expression in mammalian systems, bacteria...selectable marker, and whether the plasmid contains Cas9. ID Plasmid Expression System Promoter Guide RNA Type...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Drosophila ID Plasmid Gene/Insert...RNA-H1-nick sgRNA-mCherry Mammalian, AAV hU6 + H1 pegRNA + nicking sgRNA No mCherry Hyongbum Kim 177180...pDAS12069_U6-pegRNA-mCherry Mammalian hU6 pegRNA BpiI for pegRNA, Esp31 for PBS-RT No mCherry Ervin Welker 177181...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...

...pAAV-CaMKIIa-mCherry CamKIIa mCherry Control Deisseroth 114470 pAAV-Ef1a-mCherry EF1a mCherry Control Deisseroth...pAAV-hSyn-DIO-hM3D(Gq)-mCherry Syn Activator, Cre-dependent DREADD Roth 44362 pAAV-hSyn-DIO-hM4D(Gi)-mCherry Syn Inhibitor...pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 Syn Cre-EGFP expression Cre Wilson 105550 pAAV.GFAP.Cre.WPRE.hGH GFAP Cre-expression Cre Wilson 107738...preparations were produced with the pUCmini-iCAP-PHP.eB plasmid (Addgene #103005) . Note on PHP.eB tropism and ...Activator C1V1 Dimidschstein 135634 pAAV-S5E2-ChR2-mCherry E2 regulatory element Activator ChR2 Dimidschstein...107738 pAAV-hSyn-Cre-P2A-dTomato Syn Cre-expression Cre Larsen 140135 pAAV-EF1a-iCreV EF1a light-inducible...rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA minBG iCre expression iCre Ting 199775 AiP13778 - pAAV-AiE0450h-minBG-iCre...

...Addgene's Viral Plasmids and Resources page for more information. Plasmids Expressing Spike and Spike ...Accessory Plasmids Envelope and Packaging Plasmids Popular Lentiviral Envelope and Packaging Plasmids Popular...Envelope and Packaging Plasmids Reporter Plasmids A few examples of reporter plasmids that can be used for...circulating variants. The table below lists expression plasmids that are suitable for generating pseudotyped...reporter vector expressing firefly luciferase. pCMV-FLuc - Retroviral reporter vector expressing firefly luciferase...Lentiviral dual reporter vector expressing firefly luciferase and ZsGreen. NL4-3 mCherry Luciferase - Lentiviral...Lentiviral dual reporter vector expressing mCherry and firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST...

... and Packaging Plasmids Common γ-retroviral envelope and packaging plasmids. ID Plasmid Type Description...with packaging plasmid Weinberg 12259 pMD2.G Envelope VSV-G-expressing envelope plasmid Trono 1733 pCI-VSVG...Ecotropic MLV expressing envelope plasmid Nienhuis and Salmon 35616 pCAG-VSVG Envelope VSV-G-expressing envelope...envelope plasmid Nienhuis and Salmon 22501 pHEF-VSVG Envelope VSV-G-expressing envelope plasmid Kasparov Popular...γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want a retroviral plasmid with your gene...10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression; also expresses GFP. See pMKO.1 puro for Puro... to eGFP expression Clevers 18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression; also expresses...

...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. Resources...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...