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We narrowed to 278 results for: guide rna expression plasmid

Showing: 121 - 140 of 278 results

  1. Bikard Lab - CRISPR Repression Collection

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    Collection
    ... the RNA polymerase, thus strongly decreasing the expression level. Mismatches between the guide RNA and...range of expression levels. This collection comprises 20 plasmids, each carrying two CRISPR guides. The first... can be controlled using a plasmid‐borne CRISPR array coding for guide RNAs (diamonds) interspaced with... CRISPR Repression Plasmids You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR...From the Bikard Lab, CRISPR plasmid collection for modulating expression levels of two targets independently...arrays to modulate the expression levels of two targets independently. The RNA-programmed DNA-binding ...nucleotides. Together, these plasmids allow to explore the whole range of expression levels for these two reporters...

  2. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...resources from Addgene Plasmids 101: Modular Cloning Addgene's CRISPR Guide Plasmids 101: Golden Gate Cloning...restriction digest, and then plasmid sequencing if you can. To quote Doench, “Whole plasmid sequencing is just ...correctly banded clones for plasmid sequencing and found 98% of the plasmids were perfectly correct. Because...pRDA_757 (Piggybac; constitutive expression) Mammalian Plasmid pRDA_791 (contains only orio and...the assembled plasmid. It shows you each component, with the ID of the component plasmid, component name...the component plasmids on the Addgene website. Step 4: Naming your assembled plasmid and generating...structure takes a lot of the design work out of the plasmid creation process. Each module of Fragmid represents...

  3. Plasmids 101: Biotinylation

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    Blog Post
    ...more on this below) it can be incubated or co-expressed with avidin or a structurally similar protein ...ABC and IHC check out Thermo Fisher Scientific's guide. There are an array of other molecular biology applications... been shown to effectively biotinylate AviTag expressing proteins in bacteria, yeast, insect and mammalian...biotinylating proteins with BirA. You can find plasmids that contain both AviTag and BirA at Addgene! ...Photobiotin can be used to non-selectively label DNA and RNA. Photobiotin acetate consists of a biotin molecule...., & Zhu, Y. “A non-radioactive method for small RNA detection by northern blotting.” Rice 7.1 (2014):... shift assay to examine specific dsDNA, ssDNA or RNA–protein interactions.” Nucleic Acids Research, 23.18...

  4. Typing CRISPR Systems

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    Blog Post
    ... binds the guide RNA (shown as mature crRNA) and binds the RNA target before initiating RNA cleavage. ...Schematic of a Cascade complex (blue subunits) with a guide RNA (green and orange) bound to the target DNA with... exclusively targets RNA. Cas13 has become a natural successor to traditional RNA interference, as it ...target and degrade select RNA transcripts, or be engineered to conduct direct RNA editing. There are four...Additional resources on Addgene.org CRISPR guide CRISPR plasmids CRISPR references ... you need to ensure every unit of the complex expresses at similar levels. It was only recently that researchers... researchers began building optimal plasmids to harness the power of Class 1 systems. Class 1 CRISPR systems...

  5. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    Blog Post
    ... one, Ly6a, showed differential protein and RNA expression between the brains of C57BL/6J mice and BALB...Deverman Lab when they looked at LY6A protein and RNA expression patterns, the PHP.B permissive phenotype of...phenotypic effects such as those that affected gene expression, mRNA splicing, or protein coding regions. In total...strong linkage to the PHP.B phenotype and its expression pattern in the mouse brain.  Both labs then took...Deverman lab started by seeing how changes in LY6A’s expression affected PHP.B’s transduction rates. Knock out...still transduced by PHP.B but only if Ly6a was expressed. Additionally when AAVR knockout mice were IV ...permissive and non-permissive mouse strains. PHP.B plasmids and PHP.eB viral preps are available from Addgene...

  6. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

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    Blog Post
    ... Visit Our Stem Cell Page Find Plasmids for Reprogramming Find Plasmids for Differentiation ...reprogramming differentiated adult cells by the ectopic expression of four embryonic genes- OCT4, SOX2, KLF4, and...to a Nobel Prize for Dr. Yamanaka in 2012. Find plasmids for generating iPSCs iPSCs and ESCs closely resemble...possess similar morphologies, growth properties, and express stem cell markers similar to ESCs [11].  While ...Pharmacological Sciences, a Cell Press reviews journal. Stem cells are special types of cells that can...Pharmacological Sciences, a Cell Press reviews journal. She joined Cell Press to pursue a career in science...

  7. CRISPR Guide

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    ...Science Guides CRISPR Guide CRISPR Guide Jump To: CRISPR Guide Overview CRISPR Basics Small Edits Large...that target RNA instead of DNA enables tracking of RNA in live cells, including orthogonal RNA/DNA labeling...) and editing (B) with Cas13 Browse Plasmids: RNA Targeting , RNA Editing Degrading DNA with Cas3 Rather...Repurposing CRISPR as an RNA-Guided platform for Sequence-Specific control of gene expression. Cell . 152 (5),...components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme). The gRNA is a short...conversions using a prime editing guide RNA ( pegRNA ). Like sgRNA, pegRNA directs the nickase to the target...delivering multiple gRNAs using a single plasmid ensures that all gRNAs will be expressed in the same cell...

  8. Overcoming the Challenges of Lentiviral Production

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    Blog Post
    ... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection... titer virus for transfer plasmids with small inserts (such as CRISPR guides) while a single 72h harvest...Resources on Addgene.org Lentiviral Guide Find lentivirus plasmids Browse all viral vectors ... a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find...

  9. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

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    Blog Post
    ...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no proteins...involved in RNA based surveillance of repetitive gene families and provide a way to guide inheritance ...finding in the regulatory RNA field. The Rajewsky lab is studying circular RNAs (who knew…I had barely heard...stranded, covalently closed RNAs seem to be conserved in mammals, expressed in neurons, and are especially...against both DNA and RNA viruses - 2 cas systems regulate at the DNA level and 1 at the RNA level. Udi Qimron...understanding complex regulation of and by tRNAs and tRNA-derived RNAs. Stay tuned for big developments in this... discover circRNAs in RNA-seq data. As far as I can tell, we don’t really know what circRNAs do yet or...

  10. 22 Hot Plasmid Technologies from 2014

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    Blog Post
    ...Cre-based Brainbow-expressing cells. Autobow plasmids are all-in-one versions which express self-excising ... host cells in order to optimize expression. The expression plasmids for a given species are based on ... Hot Plasmid list. Other popular plasmids & blog posts @Addgene: The 10 Most Distributed Plasmid Technologies...inducible inactivation. Find the Robinson plasmids or the Royle plasmids and get to work! Robinson et al...descriptions of various optogenetics plasmid tools, visit Addgene’s optogenetics guide. More optogenetics tools...designed the pOSIP (one-step integration plasmid) series of plasmids, vectors that convey both the sequence...Chen, pCS2-3xFLAG-NLS-SpCas9-NLS is a Cas9 expression plasmid that was used by Chen and colleagues for ...

  11. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

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    ...lentiCRISPRv2, expressing both Cas9 and an individual gRNA from this single vector) and guide-only (lentiGuide...gRNA plasmids targeting the human kinome. These plasmids contain the same 3-4 gRNAs found in the first ...setting. The individual plasmids will also be useful for imaging or gene expression assays necessitating ...lentiGuide expressing only an gRNA, for use with cells that already have Cas9). Each library consists of ...individual plasmid formats A first for a library at Addgene, you can also order the individual lentiGuide gRNA...individual plasmids separately. Additionally, this set can be used for cherry-picking individual gRNAs that ...Pooled Library Page Find Validated gRNAs Deposit Your CRISPR Plasmids ...

  12. Plan Your Experiment

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    Collection
    ...Cas enzyme and guide RNA) for your experiment (Figure 1). You will decide how to express Cas9, the delivery...delivery system for Cas9 and the guide RNA (gRNA), the design of the gRNA sequence, and finally how to validate...used for gRNA design. Addgene offers many plasmids containing validated gRNAs . These plasmids contain ...and point mutations using a prime editing guide RNA (pegRNA) Prime editor (Cas9 nickase fused to reverse...is to directly transfect expression plasmids encoding your Cas protein and gRNA, which you generally perform...off-target effects. RNA delivery The first of these methods is RNA delivery of Cas and gRNA. This method involves... Science Guides CRISPR Guide Plan Your...

  13. Lentivirus Production

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    Protocol
    ...DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your insert of interest...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be... 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios...

  14. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...with the guide RNA.   Non-target strand The DNA strand that does not base pair with the guide RNA. Despite...Acronym Full name Notes CRISPR terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching DNA ...structure. In order to bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and (3) a protospacer-adjacent... called an R-loop (Figure 2). This allows the guide RNA to base pair with the DNA target strand, leaving...binding to DNA and separating the two strands. The guide RNA base pairs to the target strand. The complementary...Addgene.org Browse CRISPR plasmids for base editing Get a broad overview with our CRISPR Guide Check out our CRISPR... reduce its tendency to make off-target edits to RNA. Like other ABEs, the editor they were using consisted...

  15. Viral Vectors 101: Pseudotyping

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    Blog Post
    ...three plasmids: (1) The transfer plasmid, (2) the envelope plasmid, and (3) the packaging plasmid. These...pseudotyped protein is encoded on the envelope plasmid which contains a promoter, the envelope gene, and...virus, HIV-2, and human foamy virus in cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars...than viruses produced in cell lines that do not express these sugars. (Takeuchi et al., 1997). The alphaGal...scientists encode the spike protein on the envelope plasmid. Image from Crawford et al., 2020.   Broad tropism...infecting most cells, but not all. For example, LDLR expression in hematopoietic stem cells, T cells, and B cells...pseudotyped virus requires the upregulation of LDLR expression using other methods.  In terms of preparation...

  16. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    Blog Post
    ...Check out our Optogenetics Guide or browse our collections of optogenetics plasmids – and keep an eye out ...resources on Addgene.org Optogenetics Guide Optogenetics Plasmids References Aljiboury, A. A., Ingram, ...clusters under blue light Check out our Optogenetics Guide to learn more about how these modules work, and ... of katanin to that anchor. So, you’ll need to express both the EB3N-VVDfast-iLID anchor module and the...module, which are also available as lentiviral plasmids if you want to make a stable cell line. This design...specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID along with your...Opto-RhoGEF design and activity. B) OptoTIAM (magenta) expressed in a monolayer of endothelial cells, showing cell...

  17. RNA Extraction Without A Kit

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    Blog Post
    ...DNA or RNA purification to avoid contamination, take extra care when performing RNA extraction. RNA is inherently...cover the basics of isolating RNA without a kit. Tips for working with RNA (whether you are using a kit... freezer to prevent RNA degradation. They will degrade in a -20°C freezer.   RNA extraction methods evolved...activity upon cell lysis, RNA is degraded. To effectively isolate intact RNA, a fast, strong protein denaturant...separate the intact RNA from DNA and proteins. Although very effective at isolating total RNA, this method requires...and RNA can then be isopropanol precipitated. This method reduced the time it took to isolate RNA from...precipitate the RNA using lithium chloride. LiCl solutions selectively precipitate RNA, but not DNA and...

  18. Viral Vectors

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    ...find plasmids that can be used to engineer various types of viruses. Lentivirus Plasmids | Guide Lentiviruses... high but transient transgene expression. γ-Retrovirus Plasmids | Guide γ-retroviruses, commonly referred...enabling long term transgene expression. Overview of Viral Plasmids While plasmids are typically used to encode...Viral Plasmids and Resources You May Also Like... Addgene's Viral Service Biosafety Guide Blog: Plasmids...eBook Overview of Viral Plasmids Which Type of Virus Should I Use? Use this guide to learn more about the...for select plasmids are now available through Addgene, learn more here . Browse Viral Plasmids by Type Visit...host genome. Adeno-associated Virus (AAV) Plasmids | Guide Adeno-associated viruses (AAVs) are commonly used...

  19. Generating Mouse Models Using CRISPR/Cas9

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    ... Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for Mouse Targeting...CRISPR/Cas system makes use of 3 components, a guide RNA (gRNA) of about 125 nt that specifies the target,...online to guide you in gRNA design. We have been using Benchling, but there are many other gRNA design tools... create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components are brought together...John Doench’s gRNA design blog post. Finally, always remember that you are working with RNAs, which are ...the target site, and a donor oligonucleotide or plasmid as the repair material if needed (for knock in ...provided (in the form of an oligonucleotide or plasmid), precise changes can be made in the genome via...

  20. The AAVantages of AAV in CRISPR Screens

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    ...addgene.org CRISPR pooled libraries Guide to pooled libraries CRISPR guide AAV guide ... to use a multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9, while the ...Addgene. The large packaging capacity and stable expression of lentiviral vectors make them an attractive...attractive option. However, safety requirements and expression limitations may be too constraining for some experiments...improved safety profile, customization, and enhanced expression in many mouse tissues. Safety AAV and lentivirus...significant degree using different types of AAV. Expression Another advantage of AAV vectors is how the cargo...cargo — in this case, the CRISPR components — are expressed. Unlike lentivirus, cargo delivered by AAV does...
Showing: 121 - 140 of 278 results