...more stable in human plasma than the standard spCas9 (Harrington et al. 2017). Lastly, as with any scientific...commonly performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains...chosen for both Cas9 and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target...targets with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for... as CMV or EF1A promoters found in many CRISPR plasmids) are not expressed. Because this method does ...fluorophore tagged dCas9 may require the use of plasmid or viral-mediated delivery. You’ve decided to...and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier’s Lab...

...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome...MORF Collection) and an accompanying expression atlas, including over 3,500 human TF isoforms, to help...understand this regulatory network. The transcription atlas and library The transcription factor library The...determine how its presence influences cell fates. The atlas of directed differentiation To understand how each... fates. (Adapted from Joung et al.)    This atlas linked TF expression to differentiation outcomes...characterized TFs, the Zhang group were able to classify unknown, or ‘orphan’, TFs into their respective...cell types generated by the human fetal expression atlas. Computational analysis was used to infer differentiation...

... 1998). Interested in plasmids that contain the groucho gene? Check out plasmids deposited by the Shivdasani... several plasmids that contain SHH, you can find them all here! The most popular SHH plasmid however was... to see for yourself. We have one plasmid in our repository, plasmid 39624, that contains cmr.   tinman...website for plasmids that contain tinman or the mammalian homolog Nkx2.5. We also have one plasmid, Addgene...Drosophila Read our Plasmids 101 blog series Resources on Addgene.org Find plasmids for your research ...the stories behind the names. We also have many plasmids in our repository that you can use to study these... Beauty (Ivics et al., 1997). We have a lot of plasmids that contain components of the SB transposon system...

...lentiviral plasmids include popular envelope and packaging plasmids as well as transfer plasmids. Download...Otherwise, check out our plasmid and DNA backgrounds. Download the blue plasmid background.     You can... the plasmids that Addgene offers by visiting our collections page. Here, you can browse plasmids by field...your favorite reagent, scientific technique, or plasmid repository at your next meeting.     If you’re...cute mascots!  Blugene has been helping us make Plasmids 101 videos and needed a break so they're hanging...coronavirus collection page, where you can find plasmid tools to accelerate your COVID-19 research.  Download...background.         If you’re predominantly a plasmid user, instead, don’t fret; we have backgrounds ...

...CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR plasmid tools...edits in protoplasts or plants of rice and wheat. Read the paper in Genome Biology Find plasmids containing...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ...paper in Nature Communications Find the Cas12b plasmids  Plant base editing with a Cas9-adenosine deaminase...Neuroscience Find the AAV-based SaCas9 genome editing plasmids String assembly gRNA cloning (STAgR) String ...the paper in PLoS One Find the STAgR cloning plasmids Mobile-CRISPRi allows CRISPRi experiments in...Gross’s lab recently deposited “Mobile-CRISPRi” plasmids, a set of modular CRISPRi tools that can be used...

...CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids Topics: CRISPR, Other CRISPR... where we highlight a few of the newest CRISPR plasmids available at Addgene. If you want to read more...will cover: Miniature CRISPR-Cas systems SEND plasmids Prime editing improvements Miniature CRISPR-Cas...Read the paper in Molecular Cell Find the CasMINI plasmids Efficient CRISPR editing with a hypercompact ...paper in Nature Biotechnology Find the Cas12f-GE plasmids Programmed genome editing by a miniature CRISPR-Cas12f...cells using various delivery methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in... in Nature Chemical Biology Find the AsCas12f1 plasmids   Selective endogenous encapsidation for cellular...

... As reviewed in our Plasmids 101 series, proteins are overexpressed from a plasmid construct, most often...multiplexing. pCri plasmids are divided into two sets, each of which includes plasmids with different varieties...on the Addgene Blog Plasmids 101: E. coli strains for protein expression Plasmids 101: Protein tags ...varieties of tags, both N- and C-terminal. Each set of plasmids uses the same restriction enzymes for cloning ...for one protein quickly and easily. These tagged plasmids also include cleavage sites should one need to...other tags used in the pCri system, please see Plasmids 101: Protein Tags. Beyond E. coli: purifying ...subtilis and Pichia pastoris pCri also includes plasmids designed for protein expression in systems other...

... by 10% plasma, but GeoCas9 remained active in up to 30% plasma. Figure 2: Human plasma longevity...activity in HEK293’s. GeoCas9 stability in human plasma The last trick GeoCas9 has up its sleeve is its increased...diluted human plasma. GeoCas9 remains active after being incubated in up to 30% human plasma. The discovery...incubated SpCas9 and GeoCas9 RNPs in diluted human plasma for 8 hours at 37°C. Substrate was then added and... SpCas9 or GeoCas9 RNPs were incubated in human plasma diluted with reaction buffer for 8 hours at either...excited about? Thermostability? Longevity in human plasma? Something we haven’t thought of yet? Let us know...you want to give GeoCas9 a try, as always, the plasmids are available from Addgene. Happy CRISPRing! ...

...three plasmids — a transfer plasmid containing your gene of interest plus two packaging plasmids (three...baculovirus plasmid. Production of baculovirus requires only two plasmids — your transfer plasmid and the ...three packaging plasmids for 3rd generation lentivirus production). The packaging plasmids are widely available...to worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities...called a bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available, so the amount ...insect and mammalian systems. Baculovirus transfer plasmids have a much larger packaging capacity — up to ...can produce AAV and lentivirus if needed # of plasmids Three or four Two Packaging limit ~4.7 kb (AAV...

...features was our collection of popular plasmid sequences. These plasmids contain features such as antibiotic... our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality control process...out full plasmid sequencing at Addgene Additional Resources on Addgene.org Example plasmid analysis ...design was done painstakingly by hand. Records of plasmid construction were often incomplete or nonexistent...popular aspects - its ability to annotate a raw plasmid sequence and display frequently used genes and ...when the DNA sequence has changed. Identifying plasmid control elements Coding sequence features are relatively... boundaries for control elements in commercial plasmids were inconsistent and sometimes clearly wrong....