...lentivirus can be used for a variety of downstream applications such as stable-cell line generation. Workflow...mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 12–...

...single assay. Overview of Antibody Applications Antibody-based applications can be generally classed into ...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...production and storage techniques, and explain common applications. Science... Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods...of other antibodies. This has useful research applications, and is covered in the following Isotypes section...signaling molecules and are often used in clinical applications. Diabodies - Diabodies contain two Fab fragments... molecules that can be distinguished in your application. The indirect detection method uses an unconjugated...

...amplify and design primers that will bind to and replicate it. The following image shows the ends of the ...mutation than restriction enzyme based cloning. DNA replication by PCR has error rates that range from roughly...

...BSA standard that range from 0–2000 µg/mL . In duplicate, dilute 10 µL of standard, blank, and lysate samples... nm . Calculate the average absorbance of the duplicate samples on the plate. Subtract the average absorbance...

...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse CRISPR Plasmids By Function Genome Editing...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted...Plasmid Gene/Insert Promoter Selectable Marker Publication Prime Edit Cas9 H840A nickase fused to a reverse...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Activate Catalytically dead dCas9 fused to a ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Interfere Catalytically dead dCas9, or dCas9 ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Epigenetics To make targeted epigenetic modifications...

...Selectable Marker PI Publication Bacteria Plasmid Gene/Insert Promoter PI Publication Drosophila Plasmid... Marker PI Publication Plant Plasmid Gene/Insert Promoter Selectable Marker PI Publication C. elegans ... Marker PI Publication Yeast Plasmid Gene/Insert Promoter Selectable Marker PI Publication Zebrafish Plasmid...Marker PI Publication Xenopus Plasmid Gene/Insert Promoter Selectable Marker PI Publication Parasites ...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Do you have suggestions for other plasmids that...

... any application Balanced Salt Solution (BSS) An ophthalmic solution suitable for any application TMN200...each of the above serotypes and buffers. Please click on the plasmid name for ordering information. ID...When using the AAV2(Y444F) serotype in future publications, please acknowledge Arun Srivastava and cite...When using the AAV2(trpYF) serotype in future publications, please acknowledge Arun Srivastava and cite... using the AAV2(4pMut)dHS serotype in future publications, please acknowledge Shannon Boye and cite Boye...using the AAV6(dbY-F+T-V) serotype in future publications, please acknowledge Arun Srivastava and cite...Strategies for Overcoming Donor-Variation and Implications in Genome Editing. Sci Rep . Oct 19;6:35495....

... the original publication below. You can also download the full-text of the publication here . Article...bands from biallelic deletion clones with a proofreading polymerase and clone the amplicons ( e.g., with... of Visualized Experiments (JoVE). The video publication by Stuart Orkin's and Daniel Bauer's labs details...single-site small indels given the efficiency of biallelic modification, ease of rapid identification by ... sgRNA located within exons will enrich even monoallelic deletion clones for loss of function. This is...useful to characterize the non-deleted allele in monoallelic deletion clones. For small deletions (as the ...is critical that the deletion and non-deletion amplicons be easily resolved on an agarose gel for multiplexing...

...Experiment Web References PAM Sequences Glossary Publications CRISPR Overview Bacteria have an interesting... every genomic target. Due to its comparative simplicity and adaptability, CRISPR rapidly became the most... to various Cas enzymes have extended CRISPR applications to increasingly complex functions, including...understanding of CRISPR biology, introduce the various applications of CRISPR, and help you get started using CRISPR... Cas12a, which are available for specialized applications. Check out our list of additional Cas proteins...genomic edits carried out by Cas9. Multiplexing applications include editing multiple genes at once; using...to help you select an optimized gRNA for your application (see: Plan Your Experiment ). In addition to ...

...efficiency than NHEJ. Plasmid Gene/Insert Promoter PI Publication Browse CRISPR Plasmids By Function Genome Editing...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...opposite strand) mutations. Plasmid Promoter PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted...repair (HDR). Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase fused to a reverse...RT template. Plasmid Gene/Insert Promoter PI Publication Activate Catalytically dead dCas9 fused to a ...specific locus. Plasmid Gene/Insert Promoter PI Publication Interfere Catalytically dead dCas9, or dCas9 ...specific locus. Plasmid Gene/Insert Promoter PI Publication RNA Targeting Type VI CRISPR systems, including...

...headings. Click on the publication link to view all plasmids available from the article. Click on the PI...headings. Click on the publication link to view all other plasmids from that article. Click on the PI...Plasmid Description Gene/Insert Vector Type PI Publication Back to Top Shuttle Vectors Search the table ...Description Vector Type Selectable Marker PI Publication Back to Top Do you have suggestions for other...

...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...library page. Find Pooled Libraries of Interest Click on different properties to create a custom filtered...Activation Human Zhang 3rd 10 96,458 Human CRISPR Metabolic Gene Knockout Library 110066 Knockout Human Sabatini...Knockout Human Olzmann 3rd ~10 13,920 Human lncRNA Splicing-targeting CRISPR Library 119977 Knockout Human... Mouse Birsoy 3rd 7–10 2,105 630 Mouse CRISPR Metabolic Gene Knockout Library 160129 Knockout Mouse Birsoy...Knockout Mouse X.S. Liu 3rd 5 5 10 49,252 Human Metabolic Gene CRISPRa sgRNA Library 187080 Activation Human... Human Birsoy 3rd Varies 32,460 Mouse Metabolic Gene CRISPRa sgRNA Library 187081 Activation Mouse Birsoy...

...Selectable Marker PI Publication Bacteria Plasmid Gene/Insert Promoter PI Publication Drosophila Plasmid...Promoter PI Publication Plant Plasmid Gene/Insert Promoter Selectable Marker PI Publication C. elegans ...Promoter PI Publication Yeast Plasmid Gene/Insert Promoter Selectable Marker PI Publication Do you have...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...

...therapeutic applications. Despite the ethical controversies surrounding non-research applications, it’s clear...just as useful (if not more so) for research applications, eclipsing past genome engineering technologies...for CRISPR genomic editing. Genome Engineering Applications In 2012, Jinek et al. first demonstrated that... PubMed lists more than 6,300 CRISPR-related publications, many of which detail work to improve the tool...various species, as well as the development of new applications. The first CRISPR papers described two main ...RNA, making it a very flexible system. Other Applications Scientists have also used the targeting capability...Biotechnology companies are exploring therapeutic applications of CRISPR to treat genetic disease, with the...

...Marker PI Publication Bacteria Plasmid Gene/Insert Promoter Selectable Marker PI Publication Plant Plasmid... Marker PI Publication Yeast Plasmid Gene/Insert Promoter Selectable Marker PI Publication Zebrafish Plasmid...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...and are thus well suited to directed evolution applications. Examples of these base editing systems include...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Do you have suggestions for other plasmids that...

...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Base Edit Catalytically dead dCas9 fused to a...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted...Description Gene/Insert Promoter Selectable Marker PI Publication Activate Catalytically dead dCas9 fused to a ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Interfere Catalytically dead dCas9, or dCas9 ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression...

... the cell and regulates the switch from anabolic to catabolic processes. Active mTORC1 promotes protein...mTOR pathway. enhanced signaling of mTOR, a key metabolic regulator, correlates with poor prognosis in many...ammalian t arget o f r apamycin (mTOR) is a key metabolic regulator controlling cell growth and proliferation...energy-intensive processes. mTORC1 also blocks catabolic processes including autophagy and lysosome biogenesis...survival and proliferation. mTOR Pathway Plasmids Click on a name to find available plasmids for the gene...Sabatini . Return to top mTOR Pathway - Gene List Click on a name to find available plasmids for the gene...

...control). The end result of the meeting will be a publicly shared practical guide for all scientists in the...useful to help improve this technology or its application? What are some current obstacles that researchers...prepare and compile resources for this meeting's publication, we are asking participants to share practical...antibodies and recommended dilutions, etc.), novel applications, or any other useful information. Experimental...have final approval before anything is released publicly, but please only share materials that you are ...materials that you have permission to share (e.g., publication's permission). Connie Cepko, Harvard Medical School...section for how to best help contribute to this publication. Citation information for openly shared communication...

...gRNA Delivery of gRNA & Cas9 Validate Results Publications Get Started CRISPR is a powerful system that...may not be a perfect plasmid for your specific application, and in such cases, it may be necessary to customize...reagent to suit your needs. Genetic Manipulation Application Cas9 gRNA Additional Considerations Knockout ...method Expression System Components of System Application Mammalian expression vector Cas enzyme promoter...gene you target will depend on your specific application. For example: To activate or repress a target...region is removed from the mRNA due to alternative splicing. Exons near the N-terminus are targeted since ...validate your edit will depend upon your specific application. However, there are several common ways to verify...

...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...NHEJ. Plasmid Gene/Insert Selectable Marker PI Publication Base Edit Catalytically dead dCas9 fused to a...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick CRISPR/Cas nickase mutants introduce gRNA-targeted...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Prime Edit Cas9 H840A nickase fused to a reverse...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Activate Catalytically dead dCas9 fused to a ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Interfere Catalytically dead dCas9, or dCas9 ...Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression...