...techniques for deletion, identification, and characterization. This strategy relies on cellular delivery... examine for scarring. This may be useful to characterize the non-deleted allele in monoallelic deletion...μl. This facilitates screening by allowing a multichannel pipette to be used for the remainder of the ...in each well) to a 96-well PCR plate using a multichannel pipette. Centrifuge PCR plate at 400 x g for...Validation of Biallelic Deletion Clones In order to characterize obtained clones and validate a successful knockout...and subject each clone to Sanger sequencing to characterize each deletion allele 5–7 . Repeating the PCR...Zhang F. Nature Protocols . 2013. PubMed . Characterization of genomic deletion efficiency mediated by...

... protein 35 ARHGEF2 Rho/Rac guanine nucleotide exchange factor 2 CCND CCND1 CCND2 CCND3 Cyclin D CDK CDK4...MLST8 MTOR associated protein, LST8 homolog MTOR Mechanistic target of rapamycin NF1 Neurofibromin 1 NFE2L2...Phosphatidylinositol-3,4,5-trisphosphate-dependent Rac exchange factor 2 PRKA PRKAA1 PRKAA2 PRKAB1 PRKAB2 PRKAG1... RAPGEF RAPGEF1 RAPGEF2 Rap guanine nucleotide exchange factor RAS KRAS MRAS NRAS Kirsten rat sarcoma ...protein SOS SOS1 SOS2 SOS Ras/Rho guanine nucleotide exchange factor SPRED SPRED1 SPRED2 SPRED3 Sprouty related...Tuberous sclerosis VAV1 Vav guanine nucleotide exchange factor 1 YAP1 Yes associated protein 1 Return ...

...activity, like transcription. Michael Davidson FP Collection Browse the Michael Davidson comprehensive collection...protein-protein interactions or conformational changes within a protein. Biosensors Monitor small biomolecules...protein of interest resides by using a well-characterized fluorescent fusion protein. In Vivo Imaging ... and UCSF Molecular Expressions Galleries from Michael Davidson and FSU Spectra Database hosted at the...

...ideally less than 10bp away. For small (<50bp) changes, you should consider using ssDNA oligo as donor...80bp of homology arms on each side flanking the change you'd like to introduce. We use Ultramer oligo ...the Cas9/sgRNAs vector (like PX330). For large changes (>100bp insertions or deletions), we typically ...to mutate the PAM 'NGG' in the HR template, like change it to 'NGT', or 'NGA', or 'NGC' (if it's within...required for successful targeting and cleavage, changing it to'NGT' or 'NGC', in addition to mutations ...typically use PAGE purified long oligos. For large changes (>100bp insertions or deletions), the Zhang lab...

...nlsdTomato Control Fishell 213829 pAAV_BiCHATe27_dTomato_nlsdTomato BiCHATe27 dTomato/nlsdTomato Control Fishell...BiSSTe10 Activator ChR2 Fishell 213830 pAAV_BiCHATe27_ChR2_mCherry BiCHATe27 Activator ChR2 Fishell 213856 pAAV_BiVIPe4...Link opens in a new window) and was adapted from Challis et al. (2022). Text version of the AAV decision...Viviana Gradinaru and cite as follows: PHP.eB, PHP.S: Chan et al., 2017 Pubmed 28671695 (Link opens in a new...

...Pathway Plasmids mTOR Gene List Resources The m echanistic or m ammalian t arget o f r apamycin (mTOR) is...mTORC2. Background mTORC1 & mTORC2 The better-characterized mTORC1 responds to various signals, including...known as STK11; serine/threonine kinase 11 mTOR Mechanistic target of rapamycin NF1 Neurofibromin 1 PRAS40...Mitogen-activated protein kinase associated protein 1 mTOR Mechanistic target of rapamycin p53 TP53; tumor protein p53... the translation machinery in cancer. Bhat M, Robichaud N, Hulea L, Sonenberg N, Pelletier J, Topisirovic...

...also like... Ras Pathway RAS Clone Collections Richard Kahn Lab Plasmids GTPase Plasmids GAP and GEF Plasmids...family of regulatory GTPases have long been characterized as members of the RAS superfamily, though recently...regulated both upstream (by GEFs, guanine nucleotide exchange factors), and downstream (by GAPs, GTPase activating...GTPases and their GEFs and GAPs: concepts and challenges Sztul E, Chen PW, Casanova JE, Cherfils J, Dacks...content for this page was generated with the help of Richard Kahn . Browse ARF GTPase Plasmids by Gene The following...

... mutations have been characterized as gain-of-function, expanding the mechanisms by which mutant p53 is...transcriptional targets is ever-expanding; the best-characterized effects of p53 are in promoting cell cycle arrest...Thrombospondin 1 Return to top References Unravelling mechanisms of p53-mediated tumour suppression. Bieging KT...early ageing-associated phenotypes. Tyner SD, Venkatachalam S, Choi J, Jones S, Ghebranious N, Igelmann ...

...transfection or viral infection efficiency, to study changes in cell physiology, or as a reference level of ...Creation of N-terminal Firefly fusions in Drosopholia Michael Rosbash 128046 pGL3basic luciferase and renilla_polyA...87789 pPTluc Firefly Insertion of 5' promoter/enchancer regions in Drosopholia Herman Wijnen 126033 pTETRIS-cargo...investigating regions controlling transcription Pete Stecha 212933 pGL4.84(hRlucCP/Puro) Renilla Vector for...investigating regions controlling transcription Pete Stecha Return to Top Expression Constructs Luciferase ...

...injection. It was identified by John Chiorini and characterized by Shannon Boye’s lab. AAV44.9(E531D) The AAV44.9...Adeno-Associated Virus Delivered to the Anterior Chamber Results in Efficient Transduction of Trabecular... of glaucoma and their reversed cell entry characteristics. Gene Ther . 2020 Apr;27(3-4):127-142. PMID... AAV44.9-Based Vectors Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther . Jun 3;...

...tagging of genes of interest with FPs in yeast. Characterization of some of the FPs in this collection can ... Schmidt P, Goedhart J, Teusink B. In vivo characterisation of fluorescent proteins in budding yeast. ...cassettes for fluorescent protein tagging in Saccharomyces cerevisiae. Yeast. 2004 Jun;21(8):661-70. doi...antibiotic selection markers for engineering in Saccharomyces cerevisiae. Microb Cell Fact. 2013 Oct 25;12...

...Request Service is subject to Addgene’s Terms of Purchase and the applicable material transfer agreements...knowledge from the start. However, we know there’s a chance that things may not turn out as expected. In the...materials. Please be aware that additional customs charges may be required before your order will be delivered...courier will contact you directly regarding these charges, if applicable....

...amplified with primers targeting the heavy chain and/or light chain variable regions. The PCR product is purified...with the plasmid(s) encoding the heavy and light chains. Antibodies are expressed for 7 days and then purified...affinity chromatography, antibodies undergo a buffer exchange using centrifugal columns. The final formulation...

...from our repository. All purchases are subject to Addgene’s Terms of Purchase ....and CRISPR tools Fluorescent Proteins from the Michael Davidson collection Protein Labeling from New England...Packaging from Addgene Parkinson's Research from The Michael J Fox Foundation Search Use Addgene's Search to...

...to measure protein interactions or conformation changes, as well as FRET standards....intermolecular or bimolecular FRET) or (2) conformational changes within a protein where the donor and acceptor are... Standards The following plasmids have well-characterized FRET efficiency values and were developed to...Brightness standard used as positive control to characterize mTurquoise2 pmTurquoise2-T2A-Venus(L68V) Brightness...

...recombinase CAG AAV Bruchas 75268 pAAV-CRY2-CreN photo-activatable cre recombinase CAG AAV Bruchas 78398 pNXRVa-HACre...pCAG-Cre-T2A-mRuby2 Cre and mRuby2 CAG Mammalian Scharffetter-Kochanek 104124 LZF40: hSyn-Cre Cre hSyn Lentiviral...GFP-9xGGS-Cre-6xHis Cre fused to supernegatively charged GFP variant T7 Bacterial Liu 62657 pAYC iCre CAG...constructs Cre-loxP recombination is used to create a stochastic choice of expression among fluorescent proteins...

...dependent 5 Bruchas 139505 pAAV-Ef1a-DIO-PPO-Venus EF1a PPO Venus Cre dependent 8, 9 Bruchas 198510 pAAV_hSyn-PdCO-mScarlet-WPRE...our optogenetics AAV collection, including channelrhodopsins, halorhodopsins, and more. Viral... AAV retro helper plasmid from the Karpova and Schaffer labs. See the Retrograde AAV Inventory Page . ...

...can be repaired by the cell’s endogenous repair mechanisms. Cas3 belongs to the Class 1 family of CRISPR..., the most abundant type found in bacteria and archaea. While abundant in nature, Class 1 systems are ...components. Figure 1: Overview of Cascade-Cas3 mechanism. Created with BioRender.com. Browse, sort, or ...

...Viral Pseudotyping SARS-CoV-2 Pseudotyped Virus Mechanisms of SARS-CoV-2 infection remain poorly understood...This approach allows researchers to safely study mechanisms of infection and to determine methods of neutralizing...of the C-terminal 18-21 amino acids and a D614G change which is present in many circulating variants. ...

...through base excision repair, creating a C to T change (or a G to A on the opposite strand). Likewise,...guanosine by the cell, creating an A to G (or T to C) change. Adenine DNA deaminases do not exist in nature,...targeted AID-mediated mutagenesis (TAM) from the Chang lab and CRISPR-X from the Bassik lab , in which ...