...Peripheral Infrastructure Vectors and an Extended Set of Plant Parts for the Modular Cloning System.” PLOS... chromosomal integration of larger expression cassettes (Shaw et al., 2023). Those looking to express ...different promoters, terminators, enhancers, and cross-cassette interactions in gene expression, and to generate...

...AAV-containing Iodixanol solution will flow from the needle set at the 40–60% interface. Make sure that the microcentrifuge...Protocol Video Watch the protocol video below to learn setup and use an iodixanol column gradient for AAV purification...forth a few times to mix the iodixanol that has settled at the bottom of the column. We recommend concentrating...

...follows: Unseal the iBlot 2 PVDF Mini transfer stack. Set the Top Stack to one side and discard the white separator...

...optimization before the field begins to converge on a set of standard tools. Additionally, aspiring genetic...

...Place the gel mix in the autoclave and run on a setting that gets the sample to at least 121 ℃ under 20...

...selection. Use an siRNA selection tool to determine a set of top-scoring targets for your gene. For example...help guide your reaction buffer selection when setting up double digests. Digest pLKO.1 TRC-cloning vector...

...fuse. A major disadvantage is that you need to be set up to grow, transform and purify DNA from yeast. ...promoters and into backbones with different selection cassettes). Although gateway cloning is a simple and efficient...

...2023 Workflow Timeline Day 1: Transform, recover, set up overnight growth (Estimated time 2-3 hours) Transformation...

...to NHEJ knockout. Base editors can make a limited set of mutations. Repress or Interfere (CRISPRi) Reduce...

...You’ll want to select a pre-stained or unstained set of standards that covers a size range appropriate...

... early-onset, where symptoms appear between a person’s thirties and mid-sixties, or late-onset, where ...individuals develop late-onset AD. There is no clear association between a gene and late-onset AD. It is more likely... results in death within 3-5 years from symptom onset. Disease mechanisms are still poorly understood ...during or after a person's mid-sixties. The early-onset form accounts for less than 10 percent of all cases...cases of Alzheimer's disease. Of these early-onset forms, a fraction of these are due to a mutation in...such as the ε4 allele, are a risk factor for late-onset AD. Many more genes have been associated with Alzheimer's...

...SLC1A3 CMV Episodic ataxia Susan Amara 32868 SETD1A SETD1A His T7 Parkinson's Cheryl Arrowsmith 32880 FLAG-MATR3...Parkinsonism, early-onset Roger Reeves 53822 SYNJ1-170_pCSdest SYNJ1 CMV, SP6 Parkinsonism, early-onset Roger Reeves...ALS Catherine Tomasetto 104450 pEGFPC1-hVAP-B KD/MD VAPB GFP, HA CMV ALS Catherine Tomasetto 104465 hnRNPA2...SERPINI1 Halo Dementia Michael Ward 178169 SETX_Halo_C_allele SETX Halo ALS, Ataxia with neuropathy Michael...C6A/C111A SOD1 ALS Nam-Chul Ha 193056 pADC10-SETXhel SETX Flag CMV, p10 ALS, Ataxia with neuropathy Andrew...infantile onset Smita Patel 205056 SUMO-Twinkle 43-684 TWNK His Spinocerebellar ataxia, infantile onset Smita...disease Kivanc Birsoy 218838 3xFlag-eGFP-Flag-SETX SETX Flag, GFP CMV ALS, Ataxia with neuropathy Albert...

... guide RNA expression cassettes and a Cas9 nuclease/nickase expression cassette using the Golden Gate ...the expression of multipe gRNAs. Csy4 cleavable cassettes; requires expression of Csy4 (eg Plasmid 53369...gRNA module vectors, multiple gRNA expression cassettes can be assembled using Golden Gate cloning or ...polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG cassettes are assembled into a Cas9 expressing vector. Yang...to rapidly generate multiple sgRNA expression cassettes, which can be assembled into the binary CRISPR...sgRNAs Zebrafish Plasmids for assembly of gRNA cassettes using Golden Gate ligation into Tol2-based pGGDestTol2LC...

...Packaging Plasmids Transfer Plasmids Lentiviruses, a subset of retroviruses, are some of the most common and...shRNA under mouse U6 promoter; CMV-EGFP reporter cassette is included to monitor expression Parijs 11619...shRNA under mouse U6 promoter; CMV-EGFP reporter cassette is included to monitor expression Kissler 1730... plasmid with a chimeric 5’LTR, Any expression cassette (promoter and gene of interest) can be cloned ...interest, also can express shRNA (cloning H1-shRNA cassettes into the unique SnaBI site in the 3´-LTR). See...

...plasmids for CRISPR-based knock-in of EGFP-2A-PuroR cassette to the C-terminus of endogenous proteins. The ... The PITCh donor plasmid with an EGFP-2A-PuroR cassette, flanked by microhomologous sequences and gRNA...PITCh plasmids were tested by fusing EGFP-2A-PuroR cassette to a nucleolar protein, fibrillarin (FBL). To ...cells that uses PCR to generate both an expression cassette for the Cas9-programming sgRNA and HR donors for... expression vector allows for efficient marker cassette FLP-out. Plasmids can be found associated with...

...active promoter like CAG, or expressed only in a subset of cells under a more specific promoter (e.g. Rho-Cre...p252 pPr-CREM/CMV-STOP-luc CREM and CMV-STOP-luc cassette probasin Mammalian Green 8408 p255 pPr-CREM/Ins...Ins/CMV-STOP-luc CREM and CMV-STOP-luc cassette, separated by insulator probasin Mammalian Green 11543...Sauer 12168 pMB80 (R26-CreER) Cre-ERT2 with loxp cassette; Targeting vector for Rosa26 locus Mammalian Jacks...either side (often called a “lox-stop-lox” or “LSL” cassette) upstream of a gene of interest will prevent gene...activates gene of interest, removes Neo and stop cassette; Contains flanking arms for Rosa26 integration...pPNT6 Cre activates gene of interest, removes Neo cassette. Used in gene targeting. Mouse Targeting Shivdasani...

...inducible expression for Gateway cloning; selection cassette in format: PGK-rtTA-2A-puro; see article for tagged...inducible expression of a miR-shRNA; selection cassette in format: rtTA3+IRES+Neo; see article for additional...conditional cell line generation; contains expression cassette for TetR-P2A-GFP; see article for additional constructs... rtTA On Alexeyev 14901 pCL-CTIG The inducible cassette contains tTA at its 5' end and an inducible promoter...

...markers. The FP together with the protothropic cassette can be amplified with the following primers: FW...in a new window) Sheff MA, Thorn KS. Optimized cassettes for fluorescent protein tagging in Saccharomyces...Bydder SF, Zhou Y, Nielsen LK. Dual gene expression cassette vectors with antibiotic selection markers for ...

...deletion in E3 and insertion of GFP expression cassette. Bunz 179202 pAd5-B6/7 ∆E3-GFP Adenoviral A block...deletion in E1 and insertion of GFP expression cassette. Bunz 16402 pShuttle Shuttle For production of...Shuttle Modified pShuttle-CMV plasmid with Gateway cassette; See article for additional plasmids Zhang 50957...

...fusion shuttle plasmid for shuttling U6-guide cassettes to make dual guide expressing plasmids dCas9-KRAB-MeCP2...plasmids to insert a second guide within a U6-guide cassette. This process is compatible with all pX458 family...