... of Cpf1 Type II CRISPR systems based on Cas9 were thought to be the simplest CRISPR systems and the easiest...is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227...Cpf1-driven systems added another option to the CRISPR toolbox. When Cpf1 was first discovered, researchers...nt vs 100 nt.) However, the newly described non-CRISPR protein NgAgo is smaller than any of these options..., so it has the potential to eclipse any of the CRISPR nucleases in AAV-based applications. Get More Cpf1...HDR over NHEJ, so comparing HDR efficiency across CRISPR nucleases and NgAgo is an important next step for...Benjamin P., et al. "Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells." Nature Biotechnology...

...on the Addgene blog Browse our CRISPR blog posts Download our CRISPR cheat sheet Read our science communication...Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources ... we’d already discussed the idea of my giving a CRISPR presentation to their students. I didn’t think ...(which could be adapted for any topic--not just CRISPR): Who am I? Where do I work? The teachers emphasized...this area. What is genome engineering? How does CRISPR work? This section was essentially background slides...protein, plasmid, gene, etc. Though I talked about CRISPR’s molecular mechanism, I was careful to keep it ...teachers and other science communicators.  What can CRISPR do? I spent more time on this section of the presentation...

... Addgene Blog CRISPR libraries Genome-wide screens with CRISPR Lentiviral CRISPR libraries ...with small molecules/drugs. With the advent of CRISPR, genome-wide knockout screens are possible with...While the entire genome can be knocked out with CRISPR, the entirety of the small molecule realm is typically...downstream manipulations as a forward or reverse screen (CRISPR, drug, etc.), just with an added baseline component...component. Alternative screening techniques While CRISPR and drugs dominate the screening scene, there are... worth considering. RNAi screens are similar to CRISPR, but instead of knocking out the genes of interest... be lethal to cells and thus not assayable with CRISPR. Another method, popularized in yeast, are temperature...

... the intended target site. With the widespread adoption of CRISPR/Cas9 technology and rapid improvements...Resources on Addgene.org Browse CRISPR plasmids Check out our CRISPR guide pages Find ready-to-use Cas9...delivery Epitope tagging of mammalian genes using CRISPR depends on successful delivery of three reagents...using custom synthetic two-part cr/tracrRNAs for CRISPR experiments (in collaboration with Integrated DNA...tagging is scalable to 96-well format. Off-the-shelf CRISPR reagents for the knock-in experiment were pre-assembled...csRNP knock-in experiments If your lab is new to CRISPR/Cas9 experiments, there is no need to worry at ...found that 75% of our highest knock-in efficiency CRISPR reagents had crRNAs that bind to the template strand...

... IPI and the Springer lab (Hao et al., 2024). In this study, we, along with the Springer lab, reported...produced from hybridoma antibodies developed in the Springer lab. Some are rabbit immunoglobulin G1 (IgG1) ... to be developed and defined. According to the Springer lab, a highly specific function-blocking antibody...   Targeting integrins in the lab The Ha and Springer labs recently showed RGD-binding integrins demanded...higher tensions before they would enable cells to spread, resulting in cells that elongated and morphed ...required and conferred less force, leading cells to spread into symmetrical circles. While elucidating the...integrins. The Ha lab — in collaboration with the Springer lab, the Jaumouillé lab, the Waterman lab, and...

...for 1-3 days. There’s also a CRISPR for that! CRISPR interference (CRISPRi) targets a catalytically dead...little while Yes, there’s a CRISPR for that, too! CRISPR activators (CRISPRa) are transcriptional activators...approach - read more about them on our CRISPRa blog! Fig. 1 CRISPR activator and repressor mechanisms...knock out the gene for your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need to generate...double strand break. Transient introduction of CRISPRi or inducible expression (more on this below) will... Promoters Plasmid 101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose for my Experiment ...resources on the Addgene website Promoter Guide CRISPR Guide Lentiviral Guide  ...

....107804 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids. To...To find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources... Addgene The Human GlycoGene CRISPR Library is a new lentiviral CRISPR library that targets human glycosyltransferase...Fluorescent timers for studying cell cycle length The CRISPR corner New from the viral service Labeling excitatory...arrays can lead to artifacts due to overexpression. CRISPR can insert the FP gene into the target gene’s native...introducing large genetic changes in C. elegans with CRISPR can be difficult since it requires the much less...oligodeoxynucleotide donors (ssODN). So how can you use CRISPR to introduce large FP genes into C. elegans? By...

... other CRISPR/Cas systems used for endogenous protein tagging, please visit our website CRISPR/Cas Plasmids... of interest with a fluorescent protein fusion. CRISPR genome editing can help accomplish this goal. The...sequence (like GFP) can be inserted at the DSB. CRISPR has allowed scientists to tag and light up endogenous...better understand normal protein function. However, CRISPR protein tagging does have its limitations, especially...plasmid assembly toolkit called SapTrap to improve CRISPR genomic tagging in C. elegans. SapTrap offers scientists...systems, including mammalian cells. CRISPaint (CRISPR-assisted insertion tagging), developed by the Hornung...interest to promote homology directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs...

...Amplification Type Link to Protocol CRISPR Knockout Human CRISPR Knockout Pooled Library (Brunello) ...Amplification Protocol (PDF, 88.2 KB) CRISPR Knockout Human CRISPR Knockout Pooled Library (GeCKO v2) ... Protocol (DOCX, 15 KB) CRISPR Activation Human Genome-wide CRISPRa-v2 Libraries 104,540–209,080...Re-Amplification Protocol (PDF, 71 KB) CRISPR Activation Human Subpooled CRISPRa-v2 Libraries 12,285–20,145 ...Protocol (DOCX, 17 KB)   For CRISPR libraries, Genome-scale CRISPR-Cas9 knockout and transcriptional...ng Solid Joung et al. Nat Protoc. 2017 CRISPR Knockout Human Epigenetic Knockout Library ...on the Addgene blog Genome-Wide Screening Using CRISPR Preparing Pooled Libraries NGS Quality Control ...

...The natural CRISPR locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to ... invading phage pathogen. Over the past decade, CRISPR tools have leveraged such host-defense mechanisms...with only minimal payloads. DAP array  Although CRISPR-Cas12a, with abilities to process and release its...leading the edge of the precision gene editing. CRISPR-Cas9, the key component to enable efficient base...multiple gRNAs for nCas9-BE, or any other kinds of CRISPR tools, without compromising their strategic advantages...for potential in vivo applications or multiplex CRISPR screening.   Using tRNAs Luckily, a fun feature... state-of-art base editor, prime editor, or any CRISPR-based gene editors, for correcting mutations or...