...From the Bikard Lab, CRISPR plasmid collection for modulating expression levels of two targets independently...Repression Collection Bikard Lab CRISPR Repression Plasmids You may also like... CRISPR Guide CRISPR Protocols...of two targets independently. The RNA-programmed DNA-binding protein dCas9 is expressed along a CRISPR...expression levels. This collection comprises 20 plasmids, each carrying two CRISPR guides. The first one...18, or 20 matching nucleotides. Together, these plasmids allow to explore the whole range of expression...amount and stoichiometry of enzymes. The pLC97 plasmid allows for quick integration of the aTc-inducible... One-Step cloning and chromosomal integration of DNA. ACS Synthetic Biology, 2(9), 537–541. DOI: 10.1021...

...expensive) synthetic fluorophores. Add a dash of DNA with an electrical jolt and cells become perfectly... Generate Fluorescent Protein Fusions Request Plasmids from the Michael Davidson Collection Find Fluorescent...

...use the chromatin IP (ChIP) technique to identify DNA binding proteins involved in histone modification...antibody posts  Resources on Addgene.org: Find plasmids encoding recombinant antibodies at Addgene ...

...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance...Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid Calarco... need Cas9 plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription none, need Cas9 plasmid Boxem 47912...elegans none, need Cas9 plasmid Ristow Do you have suggestions for other plasmids that should be added to...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a...CRISPR plasmids that have been designed for use in C. elegans. CRISPR...

... Plasmid Collections Nuclear Receptor Plasmid Collection Nuclear Receptor Plasmid Collection...Receptor Plasmids Find Plasmids by Gene Name Alternative Gene Names Gene Description AR plasmids AR, RP11...molecules, and 2) a DNA binding domain, which targets the receptor to specific DNA sequences. Ligand binding... Nuclear Receptor plasmid collection for the community. If you have created plasmids that you would like...ESRRB Plasmids ESRRB, DFNB35, ERR2, ERRb, ESRL2, NR3B2 estrogen-related receptor beta ESRRG Plasmids ESRRG...group A, member 1 PGR Plasmids PGR, NR3C3, PR progesterone receptor PPARA Plasmids PPARA, SC22CB-5E3.5,...receptor gamma RARA Plasmids RARA, NR1B1, RAR retinoic acid receptor, alpha RORA Plasmids RORA, ROR1, ROR2...

... and Packaging Plasmids Common γ-retroviral envelope and packaging plasmids. ID Plasmid Type Description...envelope plasmid Kasparov Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want...retroviral plasmids at Addgene, including popular packaging, envelope, and transfer plasmids. Viral...Plasmids γ-Retroviral Plasmids γ-Retroviral Plasmids Additional Resources: γ-Retrovirus Guide Biosafety...Biosafety Guide Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook γ-Retrovirus (gamma-retrovirus) is an...with packaging plasmid Weinberg 12259 pMD2.G Envelope VSV-G-expressing envelope plasmid Trono 1733 pCI-VSVG...envelope plasmid Nienhuis and Salmon 35616 pCAG-VSVG Envelope VSV-G-expressing envelope plasmid Nienhuis...

... non-targeting guides. Toxicity is likely due to DNA damage and its subsequent repair. Additionally, the...on Addgene.org Check out this guide to pooled plasmid libraries Find all of the CRISPR Pooled Libraries...fail to account for the effects of Cas9-induced dsDNA breaks. Are off-target cutting patterns in genome-wide...function, i.e. sites that lack open chromatin marks, DNase hypersensitivity, or are in an enhancer, transcription...control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting vs. safe-targeting...for the inherent effects, or background noise, of dsDNA breaks caused by CRISPR. See supplemental figure...

...Cell-Imaging Plasmid Collection New England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental...Biolabs has deposited plasmids at Addgene to generate SNAP, CLIP, ACP or MCP tagged plasmids for visualization...based on the human O6-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein, and is suitable for...synthases needed for use of these tags. Plasmids Addgene ID Plasmid Substrate Feature 101137 pSNAP-tag (T7...Control Plasmid SNAP-tag Control for mitochondrial localization 101124 pSNAPf-H2B Control Plasmid SNAP-tag...Control Plasmid CLIP-tag Control for nuclear localization 101130 pCLIPf-Cox8A Control Plasmid CLIP-tag...Control Plasmid ACP/MCP-tag Control for cell surface localization 101131 pACP-GPI Control Plasmid ACP/MCP-tag...

...Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Introduction This protocol... plate and select transformed cells containing plasmids differing in their resistance genes, as one does...or other antibiotic) E. coli transformed with a plasmid containing the carbenicillin (amp) resistance gene...effective at selecting only bacteria containing the plasmids of interest (for example, fewer satellite colonies...transform DH5α E. coli cells by heatshock with the plasmid of interest. See our transformation page for a ...

...little variation. Recombinant plasmids - Antibodies can be produced from plasmids, with an antibody’s genetic... genes from specific hybridoma cells into plasmids. Plasmids are then transfected into cells and induced...interactions between DNA and protein. Micrococcal nuclease digestion is used to shear the DNA in these assays... uses antibodies to capture protein:DNA complexes. After capture, the DNA is released from the complex...suitable for DNA:histone modifier interactions, which have strong binding between the protein and DNA. Some ...immune cells, or through the use of recombinant plasmids. Animals - Antibodies produced in animals make...genetic code inserted into the plasmid. This produces the least amount of variation between antibodies, allows...

...HDR design; Deep sequencing Cas9: pMLM3613 dsDNA donor plasmid backbones pSL1180-HR-PUbECFP & pSL1180polyUBdsRED...collaboration with the labs who have deposited CRISPR plasmids , we've created a series of links and guides to...Addgene blog . Eukaryotic Pathogen CRISPR guide RNA/DNA Design Tool (EuPaGDT) gRNA design tool with extensive...Joung Lab has been expanded to identify potential DNA target sites for TALEs and CRISPR/Cas. CRISPRdirect...on Addgene's blog Protocols Lab(s) Description Plasmids in protocol Download protocol Church gRNA design...gRNA design and cloning for Cas9 orthologs Cas9 plasmids PDF 104.1 KB Chen and Wente Zebrafish: gRNA cloning...PDF 105.8 KB Fujii iChIP/enChIP to purify genomic DNA FLAG tagged dCas9 PDF 107.4 KB Goldstein Nematode...

...Broad GPP genome-wide Brunello 73179 (1 plasmid) 73178 (2 plasmid) Knockout Human Doench and Root 3rd 4 ...76,441 Broad GPP genome-wide Brie 73632 (1 plasmid) 73633 (2 plasmid) Knockout Mouse Doench and Root 3rd 4 ...GPP kinome Brunello 75314, 75315 (1 plasmid) 75312, 75313 (2 plasmid) Knockout Human Doench and Root 3rd... 3,052 Broad GPP kinome Brie 75317 (1 plasmid) 75316 (2 plasmid) Knockout Mouse Doench and Root 3rd 4 ...20,051 Human GeCKO v2 1000000048 (1 plasmid) 1000000049 (2 plasmid) Knockout Human Zhang 3rd 6 123,411...185,836 Mouse GeCKO v2 1000000052 (1 plasmid) 1000000053 (2 plasmid) Knockout Mouse Zhang 3rd 6 130,209... - 3 plasmid system 1000000075 (Puromycin) Activation Mouse Zhang 3rd 3 69,716 SAM v2 - 2 plasmid system...

...freezer, you’ll probably find lots of different plasmids containing the same protein of interest tagged...developed a self-labeling fusion tag based on the human DNA repair protein AGT that gets covalently linked to...Addgene’s collection, so you may be able to get a plasmid ready-to-use without any cloning at all! As mentioned...

...Mammalian ID Plasmid Gene/Insert PI Publication Do you have suggestions for other plasmids that should ...maintained by the Doudna Lab . Browse, sort, or search the tables below for CRISPR plasmids for RNA editing...CRISPR plasmids based on Cas13 designed to precisely edit RNA targets. CRISPR...Plasmids: RNA Editing Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit Transcriptional...enzymes Cas13a/C2c2 and Cas13b, target RNA rather than DNA. Fusing the catalytic domain of ADAR2(E488Q) adenosine... be added to this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve...

... the mammalian brain (Wickersham et al., 2007; plasmids at Addgene). AAV vectors: A platform for delivering...neuronal cells. The AAV genome is a single-stranded DNA molecule and AAV vectors are replication deficient...involved in learning and memory (Kohara et al., 2014; plasmids at Addgene). The Gradinaru Lab at the California... throughout the brain based on its unique hue (plasmids at Addgene). A large-scale example of neuronal...than any other AAV serotype (Tervo et al., 2016; plasmid at Addgene). Its efficiency is comparable to conventional...Addgene.org Find ready-to-use AAV Preps Find AAV plasmids Browse all viral vectors ...

... Plasmid Collections Cre-lox Plasmids Cre-lox Plasmids Additional...Cre-lox Plasmids Addgene's Blog: Cre-lox Addgene's Blog: FLEx Vectors Cre-containing Plasmids loxP Constructs...circular piece of DNA (and is not maintained). Translocation: If the sites are on separate DNA molecules, a...viral preparations of plasmids that are currently in our repository. ID Plasmid Description Promoter Expression...loxP plasmids, reporter genes indicate which cells have undergone Cre recombination. These plasmids can...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail... Cre-lox plasmids for your experiments. Plasmid...

...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...CRISPR/Cas plasmids for labeling and visualization of genomic loci. CRISPR... CRISPR Visualize CRISPR Plasmids: Visualize...Visualize Genomic Loci Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit Transcriptional...researchers have turned dCas9 into a customizable DNA labeler compatible with fluorescence microscopy in... sites throughout the chromosome. Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...

...aspirate supernatant. Extract genomic DNA according to the GeneJet Genomic DNA Purification Kit instructions....Thermo Scientific, 10199-452 Reagents GeneJet Genomic DNA Purification Kit, Thermo Fisher, K0721 6-well tissue...diluted and used to transduce HEK293T cells. Genomic DNA is extracted from the target cells and assayed for...green) and negative (black) droplets Sample ID Plasmid ID Dilution Copies/20 µL RRE (FAM) Copies/20 µL...

... like... Keith Joung Lab plasmids Daniel Voytas Lab plasmids CRISPR plasmids The Joung Lab has developed...Gene Additional Plasmids compatible with the REAL Assembly TALEN Kit: TALE-LSD1 plasmids for epigenome ...arrays on a single architectural framework and the DNAs produced by all three methods are identical in sequence... simplify the construction of TALEN expression plasmids using REAL, REAL-Fast or FLASH assembly. This ...REAL or REAL-Fast assembly process or lists of plasmid ID numbers required for FLASH assembly. Links to...Assembly TALEN Kit Individual REAL TALE Repeat Plasmids TALEN Expression Vectors for REAL, REAL-Fast and...

...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... 68306 C321.ΔA.Δserb.Amp S Plasmid 68292 SepOTSλ Plasmid 68307 SupD Plasmid 34623 SepOTSα SepRS/EF-Sep...pKD-SepRS-EFSep) Plasmid 34624 SepOTSα tRNA-Sep ( aka. pCAT112TAG-SepT) Plasmid 68283 SepOTSβ Plasmid 68284 SepOTSγ...( 188537 ) is a ROP minus plasmid and is not compatible with other plasmids containing high copy ColE1...

...relatively new technology that takes advantage of DNA sequencing to turn mAb clones into expression constructs...Antibodies 101: ValidationSelecting the Right AntibodyPlasmid-based Recombinant Antibodies    ...

...Table 2. pmax dCas9 Activator Expression Plasmids ID Plasmid 48223 pAC91-pmax-dCas9VP64 : dCas9VP64 on...CRISPR Cas9 gRNA plasmids from Dr. Rudolf Jaenisch's lab. CRISPR... CRISPR Jaenisch Lab CRISPR Plasmids Jaenisch Lab...Lab CRISPR Plasmids Available from Addgene You may also like... CRISPR Guide CRISPR Protocols gRNA Design...48223~48227] + sgRNA expression vector or the linear DNA containing the sgRNA expressing fragment produced...different systems by LR clonase reaction. These plasmids are described in: Multiplexed activation of endogenous...Link opens in a new window) Article . Individual plasmids can be ordered via the links below: Table 1. dCas9...

...of the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. ... 2022.   Find CRISPR Csm plasmids here! Colognori D, Trinidad M, Doudna JA. Precise transcript targeting... missed something smokin'? You can pitch a Hot Plasmids section (antibodies and viral preps welcome!) ...inducible expression.   Find heat-inducible lentiviral plasmids here!   Wu Y, et. al. Control of the activity ...2: Addgene’s western blot for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP... Campbell et al, Nature Methods   Find YFP plasmids here!   Campbell, B.C., et al. Chemically stable...novel RNA targeting system by: Susanna Stroik The Doudna lab’s recent work characterized a new tool for ...

...in analysis Plasmid-based proteins: plasmid sequence not accurate Check plasmid sequence for frameshifts...specific compartment. If your proteins are nuclear- or DNA-binding proteins, you may need to sonicate your lysates...

...packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid which provides...going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs which are... AAV transfer plasmids. It’s easy to contaminate a workspace with plasmid, not that I would know about...prep. AAV samples are first DNaseI digested to remove any residual AAV plasmid that was carried over from...sample for the DNAseI digest. Use good sterile technique when handling your AAV plasmid standard and AAV...AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate...mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged...

...We've developed a new plasmid toolkit, the Broad Institute Target Accelerator Plasmid Collection, that we...phenotypes. Broad Institute Target Accelerator Plasmid Collections To overcome this first challenge, we...developed the Broad Institute Target Accelerator Plasmid Collections, now widely available to the scientific...in our recent publications, we used the cancer plasmid collections listed above to demonstrate how gene... hope that by releasing these new and exciting plasmid collections (together with the data generated from...patients. We hope you find these new cancer variant plasmid collections useful for your studies. Many thanks... Addgene.org Fine More Kits at Addgene Browse Plasmids from the Broad Institute ...

...you find what you need.   More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular...need. You will also have to make sure that your plasmid has been incorporated into the host organism, usually...open reading frame (ORF) of interest –  try pRS plasmid shuttle vectors. You may prefer the more traditional...details and guidance for choosing your perfect plasmid backbone. Or just ask –  leave a message in the...Molecular Cloning Guide & Other Plasmid Protocols List of common epitope tags ...mammalian expression vectors, c-Flag pcDNA3, comes from the pcDNA series. It has the CMV promoter for ...protein will allow you to visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently...

...information on shipping plasmids to your country. Curious about how rabies plasmids are used in the lab? ...If you're an international researcher who uses plasmids containing genes from rabies and vesicular stomatitis...may have noticed it’s been easier to order these plasmids. This is because in 2024 certain export control...controls prescribed by the Australia Group, for these plasmids for the majority of the countries we ship to, ...lasting basis.   Table 1: Popular Rabies and VSV-G Plasmids Name Purpose Depositing Lab pCMV-VSV-G ... Weinberg pMD2.G VSV-G envelope expressing plasmid Didier Trono pVSV-G Expresses VSV-G envelope...control exceptions for rabies and VSV-G genes, plasmids ship out approximately three weeks faster, as ...

...four different plasmids into the producer cells (two packaging plasmids, an envelope plasmid, and the lentiviral...generation systems consist of an envelope plasmid and a packaging plasmid encoding HIV proteins gag, pol, rev...generation systems require an envelope plasmid, and two packaging plasmids, one encoding gag and pol and a second...transfer plasmids replication competent? What is SIN? A6: Most (if not all) Addgene transfer plasmids are ... 30% of the transfer plasmid. Many transfer vectors are SIN. Packaging plasmid(s) contain the minimal...Addgene's most popular lentiviral plasmids Overview of Lentiviral Packaging Plasmids Lentiviral Protocols and ...collection of lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include...These plasmids contain both untagged and tagged versions of SARS-CoV-2 ORFs and include plasmids that ...scientists through plasmid sharing and more. As you’ve read above, some of these plasmids have been in the...