...reaction. Many antibodies can be purchased already conjugated, which can save significant time in the lab. However...choice, you can conjugate antibodies in the lab. The complexity of this can vary significantly based on ... B cell can clone itself to enable higher production of a single antibody, but each B cell can only produce... and can be used to increase flexibility or specificity in antibody-based methods. Antibodies can recognize...antibodies can be isotype-specific or even isotype subclass-specific. This specificity can be used to ...that you can detect in a single assay is limited by the number of signaling molecules that can be distinguished...may be present in. However, polyclonals can vary significantly from lot to lot, as immune responses have...

...proteins can be functionally expressed in bacteria under the proper conditions and, epitope tags can be used...protein. Protein purification can be challenging, but once you get it right, you can learn many things about...promoter element can be used to quickly determine whether or not a particular protein can activate expression...enzyme at a particular point in bacterial growth? Can I produce large amounts of this protein and study...addition to many questions about bacteria themselves, can be answered using model bacterial systems and plasmids... Although overexpressing or knocking down a gene can tell you much about its function and/or alter cellular... genome itself. Manipulating the genome directly can give you more subtle control over protein expression...

...viral pseudotyping" can be used to target specific cells or tissues. Overall, viruses can efficiently get ... they can be delivered to a host and generate virus (right), similar to how natural viruses can be delivered...researchers can introduce plasmids to a host to generate virus. Furthermore, these plasmids can be modified..., plasmids can be used to create viruses that encode that protein. Then, those viruses can be used to ...commonly used in research, and to find plasmids that can be used to engineer viruses for a wide array of research... by Type Visit these links to find plasmids that can be used to engineer various types of viruses. Lentivirus...because of their mild immunogencity. These viruses can direct long-term transgene expression, but generally...

...fusion; can be used for cDNA expression; puro resistance Sabatini 14883 FUGW 3rd hUbC-driven EGFP; can be ...research. Lentivirus can transduce both dividing and non-dividing cells without a significant immune response... lentiviral envelope and packaging plasmids that can be used with 2nd and 3rd generation lentivirus technologies...Description PI 12253 pRSV-Rev 3rd Packaging Contains Rev; can be used with 3rd generation transfer plasmids along...12251 pMDLg/pRRE 3rd Packaging Contains Gag and Pol; can be used with 3rd generation transfer plasmids along...psPAX2 2nd Packaging Contains Gag, Pol, Rev, and Tat; can be used with 2nd and 3rd generation transfer plasmids...R8.2 2nd Packaging Contains Gag, Pol, Rev, and Tat; can be used with 2nd and 3rd generation transfer plasmids...

...present, the virus can spread from this cell to synaptically connected cells (meaning, it can spread to cells...Thus, G-deleted rabies can infect a single "starter" cell, but new virus cannot spread to any neighboring...rabies virus-based monosynaptic tracing. These AAV can be used with modified rabies virus to perform retrograde...Serotype PI 52473 pAAV-synP-FLEX-splitTVA-EGFP-B19G Can be used to complement deletion-mutant rabies virus...pAAV-syn-FLEX-splitTVA-EGFP-tTA These AAV together can be used to complement deletion-mutant rabies virus...infected cells that lack the G protein, the virus can replicate but the newly formed viruses are unable... through complementation, G-deleted rabies virus can spread to monosynaptically connected cells. To further...

... alterations can be generated within a month. phsp70-Cas9 and a single chiRNA plasmid can be coinjected... a variety of Cas9-mediated genome modifications can be efficiently generated in Drosophila and transmitted...of phsp70-Cas9 and plasmids encoding two chiRNAs can induce defined deletions between the two cleavage...sites, while the addition of an ssODN donor template can mediate gene replacement by homologous recombination...snRNA. In both vectors, the target site sequence can be added by inserting an oligonucleotide linker into...opens in a new window) Article Individual plasmids can be ordered via the links below: ID Plasmid 46294 ...

...vector can be digested using BbsI, and a pair of annealed oligos (design is indicated below) can be cloned...plasmid can be digested using BbsI, and a pair of annealed oligos (design is indicated below) can be cloned...Protocol 237.2 KB SpCas9 Plasmids The CRISPR/Cas system can be implemented in mammalian cells by co-expressing...additional guanine nucleotide to the 5’ of 20-bp guide can increase targeting efficiency. SpCas9 and SpCas9n...cassette expressing human codon-optimized SpCas9; sgRNA can be co-transfected as PCR amplicons (Ran et al., Nature...Individual human codon-optimized SpCas9 plasmids can be ordered via the links below: 42230 : PX330; SpCas9... below. Pooled genome-wide human SAM library SAM can be combined with a human genome-wide library to activate...

...deaminase protein becomes a specific base editor that can alter DNA bases without inducing a DNA break. Base...reduce unwanted off-target effects. Nickase mutants can also be used with a repair template to introduce ...dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. Design...to a transcriptional repressor peptide like KRAB, can knock down gene expression by interfering with transcription...Type VI enzymes that function in mammalian cells can be used to attentuate RNA levels. In mammalian systems...Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA and its...Cas9 (dCas9) fused to a fluorescent protein (FP) can help visualize specific genomic loci using fluorescent...

... line that can be transfected at high efficiency Lentiviral transduction Cas9 and gRNA can be present ...components. Selecting a specific genetic manipulation can be a good way to narrow down which reagents are appropriate...efficiency compared to NHEJ knockout. Base editors can make a limited set of mutations. Repress or Interfere... Mammalian expression vector Cas enzyme promoter can be constitutive (CMV, EF1alpha, CBh) or inducible...lines Useful for difficult to transfect cell types, can also be used in vivo A common choice for conducting...component expression may decrease off-target effects Can be used to generate transgenic embryos Cas9-gRNA ...protein product is produced. Alternatively, gRNAs can be designed to target exons that code for known essential...

...plasmids can be found on the Jorgensen Lab page. More information on miniMos and MosSCI plasmids can be found...cell biology, development, and aging. Scientists can knock down genes by feeding worms bacteria expressing...table. For a more complete list of plasmids that can be used in worms, search Addgene's entire collection...Collections are plasmids grouped by a specific topic, but can only be ordered individually. CRISPR Read Addgene's...CRISPR resources, along with links to protocols, can be found below: SapTrap-SEC kit for C. elegans genome...cassette (SEC). More information regarding this kit can be found at the Goldstein Lab website . SapTrap CRISPR...expressed from pHO4d-Cas9 from Michael Nonet. Protocol can be found at the (Link opens in a new window) Seydoux...

...interactions between zinc fingers within an array can reduce their specificity. Two years ago, groups led...targets thymine, and NN targets guanine (though NN can also bind adenenine with lower specificity). With... is composed of 18 TAL effector repeats. The kit can be used to make arrays from 12-31 repeats. Perhaps... TAL effector arrays is the ease with which they can be made. Intuitively, one would expect that assembling...s group from the University of Minnesota, arrays can be assembled in a matter of days. In an open access...groups describe a set of customized plasmids that can be used with the recently developed Golden Gate cloning...Addgene , custom arrays consisting of 12-31 repeats can be assembled and inserted into a variety of backbones...

...archaebacteria, that can be opened and closed in response to light. These proteins can be expressed in specific...Archaerhodopsins and Leptosphaeria rhodopsins, are pumps that can actively move cations or anions into or out of the...expression and localization. Many other types of proteins can also be controlled by light, generally by utilizing...light-oxygen-voltage-sensing (LOV) proteins. These fusions can then be used as an optical switch to control protein...

...monocytogenes , which can inhibit the CRISPR activity of both LmoCas9 and SpCas9. Acr proteins can be used to control...desired edit can increase efficiency. Synchronizing cells or modulating the cell cycle can help as HDR .... CRISPR imaging can visualize chromatin and RNA dynamics in live cells. Labeling can be done by fusing...capabilities , as it can process multiple gRNAs under a single promoter, and can cleave ssDNA non-specifically...and quantification and can be used on lateral flow strips. Other Cas enzymes can be added to SHERLOCKv2...shredder” system that can cause large genomic deletions. The boundaries of these deletions can be defined, either... an enzymatically inactive form of Cas9; can bind, but cannot cleave DNA DSB D ouble- S trand B reak, ...

...production. You can do this whenever you are ready. Payment information must be added before we can begin processing.... It has low immunogenicity, is easy to use, and can be safely handled at the lowest biosafety protection...choices! By using AAV Packaged on Request, scientists can start experiments as soon as they receive their AAV...and volume (1 mL, 2.5 mL, or 5 mL). Currently, we can only package plasmids in our collection in the serotypes...here. Details Scale and Titer Packaged on Request can be ordered in three sizes. For all viral vector preparations... request and we’ll let you know whether the item can be packaged. Addgene does not perform functional ...

...protein becomes a specific cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine...reduce unwanted off-target effects. Nickase mutants can also be used with a repair template to introduce ...dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. Design...to a transcriptional repressor peptide like KRAB, can knock down gene expression by interfering with transcription...Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA and its...gRNA plasmids that do not co-express a Cas protein can be paired with a wide variety of Cas-containing plasmids...

...protein becomes a specific cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine...reduce unwanted off-target effects. Nickase mutants can also be used with a repair template to introduce ...dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. Design...to a transcriptional repressor peptide like KRAB, can knock down gene expression by interfering with transcription...sequence, they adopt an enzymatically active state and can bind and cleave additional RNAs regardless of homology...Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA and its...gRNA plasmids that do not co-express a Cas protein can be paired with a wide variety of Cas-containing plasmids...

...footprints, and can be scaled up for systematic editing of multiple genes. Plasmids can be found associated...details on how to use this plasmid in your experiments can be found below and in the supplemental materials ...Dalvai et al. Cell Rep. 2015 . Alternatively, cDNAs can be cloned directly into this vector and targeted ...HDR-mediated gene knock-in. The detailed procedure can be found in Sakuma et al., Nature Protocols, 2016...essential nuclear and cytoplasmic proteins, which can then be depleted very rapidly after the addition ...addition of auxin to the culture medium. Plasmids can be found associated with the following article: Natsume... for selectable genome modification. This system can be used to create C- and N-terminal epitope tags....

...relatively low immunological effects in vivo . While AAVs can be handled at BSL-1, AAVs expressing oncogenes or...or toxins should be handled at BSL-2. AAV can transduce both dividing and non-dividing cells with a low...integrate into the host genome, transgene expression can be long-lived. The utility of AAV is currently limited... a helper virus. Instead, specific gene products can be provided by helper plasmids (e.g., pHelper) and...helper plasmids for packaging your AAV product. You can search or this table based on plasmid name, capsid...required to produce infectious AAV particles. You can search or sort this table based on plasmid name, ...

...nucleases in different cell types. Zinc finger arrays can be transferred in-frame to these vectors....ZFC reagents for either OPEN or modular assembly can be easily transferred in-frame to any of these vectors... Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute for pMLM335 described in ... Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute for pMLM336 described in ...

...FAQs How can I quickly check if the cloning of my oligo into a backbone like PX330 worked? You can quickly...concept of the double nickase system is that you can express two different chimeric gRNAs with the Cas9...Cas9 cuts 3-4bp upstream of the PAM sequence. There can be some off-target DSBs using wildtype Cas9. The ...Cas9n (nickase). Homologous Recombination (HR) FAQs Can you summarize HR template vector design guidelines... shorter. What is the maximum amount of DNA that can be inserted into the genome using CRISPR/Cas for ...Any reference for using ssODN as HR Template? You can take a look at this paper (Soldner et al. , Cell.... introduction of a mutation into the genome, how can cells with that mutation be selected/screened? Before...