...could be programmed for targeted DNA cleavage in vitro. In 2013, Cong et al. and Mali et al. described ...gRNAs: Truncated gRNAs display less off-target activity. High fidelity Cas9s: Three high fidelity Cas9...eSpCas9, and HypaCas9 display very low off-target activity due to rationally designed mutations. Researchers...

...RNAs regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which...CRISPR systems with wiki entries describing enzyme activity, experimental considerations, and more, created...

...Regulate Biological Activity Use fluorescent proteins to modulate biological activity, like transcription...

...BsmBI In vitro transcription none, need Cas9 plasmid Chen and Wente 42250 DR274 T7 BsaI In vitro transcription...

... that can be used for measuring infectivity and neutralizing activity are listed below: pBOBI-FLuc - Lentiviral...

... arrays listed show transcriptional activation activity of three-fold or greater in a well-characterized...reporter system but have NOT yet been tested for activity as ZFNs in zebrafish. Target Gene Name Zinc Finger...

...lipid droplets, the nucleus, and mitochondria. The activity of GTPases is regulated both upstream (by GEFs...with each component involved in GTPase signaling activity. Figure 1: Locations of ARF superfamily members...

...regulators, increase its stability and DNA binding activity, and allow it to bind transcriptional co-activators... causes a large reduction in tumor suppression activity, leading to the development of tumors in early...

... ID Name Insert Function Antibiotic Resistance Activity PI 52962 lentiCas9-Blast Sp Cas9 Cut Blasticidin...activator . ID Name Insert Antibiotic Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none...

...CRISPR systems with wiki entries describing enzyme activity, experimental considerations, and more. CasPEDIA...104.1 KB Chen and Wente Zebrafish: gRNA cloning, in vitro transcription, injection gRNA core ; Cas9 ; optimized...

...with PCR, purity assessment by silver staining, in vitro and (when possible) in vivo testing, and full sequencing...Molecular Tools Clear Filters ID Name Promoter Activity Category PI 37825 AAV-CAG-GFP CAG GFP Control ...

...EF1a GFAP/GfaABC1D Synapsin E2 regulatory element Activity Cre-dependent Flp-dependent Cre and Flp-dependent...PHPeB Clear Filters ID Name Promoter Sensor Label Activity Serotype PI Acetylcholine Sensors 137950 pAAV....

...-4T2 Parkin RBR PRKN GST tac Alzheimer's Rachel Klevit 31606 pHA-deltaN335-Kin1A KIF5A HA CMV ALS Anthony...ALS Eugene Yeo 156883 pD649-HAsp-CSF1R-Fc(DAPA)-AviTag-6xHis CSF1R Fc(DAPA), Avi, His CMV, SP6 Hereditary...leukoencephalopathy Chris Garcia 156931 pD649-HAsp-ERBB4-Fc(DAPA)-AviTag-6xHis ERBB4 Fc(DAPA), Avi, His CMV, SP6 ALS Chris...Chris Garcia 157115 pD649-HAsp-TREM2-COMP5AP-AviTag-9xHis TREM2 COMP5AP, Avi, His CMV, SP6 Alzheimer's ...Alzheimer's Chris Garcia 157447 pD649-HAsp-CSF1R-COMP5AP-AviTag-9xHis CSF1R COMP5AP, Avi, His CMV, SP6 Hereditary...leukoencephalopathy Chris Garcia 157495 pD649-HAsp-ERBB4-COMP5AP-AviTag-9xHis ERBB4 COMP5AP, Avi, His CMV, SP6 ALS Chris...

...acid can also be used to intentionally change the activity of a protein (e.g. converting a DNA binding protein...protein to a DNA cleaving enzyme) or to regulate the activity of a protein so that it is responsive to specific...

...containing MGC138371, U2B, URP VDR vitamin D (1,25- dihydroxyvitamin D3) receptor NR1I1 VEGFA vascular ...C-X-C motif) ligand 1 (melanoma growth stimulating activity, alpha) FSP, GRO1, GROa, MGSA, MGSA-a, NAP-3, ...AUTS9, HGFR, RCCP2, c-Met MIA melanoma inhibitory activity CD-RAP MIF macrophage migration inhibitory factor...

... Arc locus Mammalian Luo 47117 pT3TS-Cre for in vitro transcription of Cre lac Bacterial Ekker 47551 pDD104...Wilson 108454 mCherry-p2A-CreERT2-FRT-kan-FRT For in vitro transcription of Cre or to recombine into BAC SP6...

...Comparing SpCas9 and GeoCas9. *For in vitro Cas9 RNP activity or longevity. GeoCas9 is active at high temperatures...Optimal Activity 35 - 45 °C 50 - 70 °C Genome Editing in Mammalian Cells? Yes Yes Longevity in Serum... from 25-80 °C. Activity is expressed as Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while... while GeoCas9 has maximum activity from 50-70 °C. The key advantage of GeoCas9 is that it functions... be incubated at high temps but still maintain activity following heat treatment. The second set of experiments...Harrington et al for more examples of GeoCas9’s editing activity in HEK293’s. GeoCas9 stability in human plasma...trick GeoCas9 has up its sleeve is its increased longevity in human serum. Harrington et al suspected GeoCas9...

...sequences, Cas12a activity gets activated. This unleashes indiscriminate ssDNA cleavage activity which cleaves...Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing) technique Sensitivity: Single molecule...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in the femto-aptomolar...SARS-CoV-2 using a CRISPR based DETECTR assay Sensitivity:10-100 copies of RNA per microliter Time: ~40... need for thermocyclers. This is followed by in vitro transcription and Cas13 is then used to detect the...visual detection of novel coronavirus SARS-CoV-2 Sensitivity: 4.6-11 copies/microliter of RNA targets in a...two crRNAs that bind to two regions improves sensitivity of the assay. This reaction also contains SSB...

...switch and turn off CRISPR activity. The ability to turn off CRISPR activity is an important step towards...thing that was missing: a way to shut off Cas’s activity. The concern is that the longer Cas remains active...edits to occur. Although methods to switch on Cas activity using light or drugs have been developed, the ... the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing editing from ~30% in control...proteins are highly diverse but most block CRISPR activity one of three ways: Inhibit DNA binding Example... Anti-CRISPR proteins (Acr's) can block CRISPR activity in many ways such as inhibiting crRNA loading,...experiments 1. Reduce off-target effects Prolonged Cas activity can increase the chance of off-target editing ...

...rapidly detect nucleic acids with high selectivity and sensitivity from patient samples. Find the DETECTR...nucleic acid detection however, either lack the sensitivity or the specificity to detect nucleic acids at...level. The Zhang lab has adapted natural RNase activity of the Cas13 protein to develop and optimize the... the method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg...CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was... your targeted nucleic acid. To increase the sensitivity of the assay, targeted DNA or RNA from a sample... SHERLOCK to detect DNA or RNA with atomolar sensitivity and single base pair mismatch specificity.  SHERLOCKv2...