l4440 empty vector feeding rnai  (Addgene inc)

Journal: Current biology : CB

Article Title: Centriole-less pericentriolar material serves as a microtubule organizing center at the base of C. elegans sensory cilia

doi: 10.1016/j.cub.2021.03.022

Figure Lengend Snippet: Key Resources Table

Article Snippet: Plasmid: L4440: RNAi empty vector , Addgene , Cat#1654.

Techniques: Recombinant, Mutagenesis, CRISPR, Sequencing, Plasmid Preparation, Software

Journal: The Journal of Biological Chemistry

Article Title: Ingestion of single guide RNAs induces gene overexpression and extends lifespan in Caenorhabditis elegans via CRISPR activation

doi: 10.1016/j.jbc.2022.102085

Figure Lengend Snippet: Components for CRISPR activation in C. elegans by bacterial delivery of sgRNAs. A , domain organization of the dCas9::VP64 fusion protein, including two nuclear localization signals (NLSs), and a FLAG- and HA-tag. B , schematic representation of the region upstream of a given transcription start site (TSS) from which promoter-specific sgRNAs are selected. C , full sequence of the two sgRNA expression cassettes in vector L4440_BioBrick-sgRNA, including BioBrick cloning sites and with individual features as indicated by color. D , Western blot analysis of total protein extracts from WT N2 nematodes versus dCas9::VP64 nematodes outcrossed x0, x2, or x4 (MIR249). The 171 kDa dCas9::VP64 protein (dC9V) is detected with a FLAG antibody in all dCas9::VP64 nematodes and an additional unspecific (unsp.) band is detected in all samples at 25 kDa. E – G , lifespan assay of WT versus dCas9::VP64 nematodes on OP50 bacteria ( E ), HT115 bacteria carrying the L4440 vector ( F ), or HT115 sgRNA scramble control (SCR) bacteria ( G ). p -values of C. elegans lifespan assays were determined by log-rank test. See Table S1 for detailed lifespan assay statistics. HA, hemagglutinin; sgRNA, single-guide RNA.

Article Snippet: For the expression of C. elegans promoter-specific sgRNAs in E . coli HT115 bacteria, the L4440 RNAi empty vector (Addgene; plasmid #1654) was modified to contain two consecutive sgRNA expression cassettes ( , ) under control of individual T7 promoters and together flanked by BioBrick cloning sites ( ) ( C ), resulting in the vector L4440_BioBrick-sgRNA.

Techniques: CRISPR, Activation Assay, Sequencing, Expressing, Plasmid Preparation, Clone Assay, Western Blot

Journal: Developmental cell

Article Title: Comprehensive endogenous tagging of basement membrane components reveals dynamic movement within the matrix scaffolding

doi: 10.1016/j.devcel.2020.05.022

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: 2017 NK2318 ten-1::LL::mNG CRISPR knock-in; qy56 This study NK2502 ddr-1::LL::mNG CRISPR knock-in; qy43 This study NK2456 ddr-2::LL::mNG CRISPR knock-in; qy44 This study NK2457 ptp-3::LL::mNG CRISPR knock-in; qy47 This study NK2477 sdn-1::LL::mNG CRISPR knock-in; qy29 This study NK2413 dgn-2::LL::mNG CRISPR knock-in; qy 82 This study NK2587 gon-1::LL::mNG CRISPR knock-in; qy45 This study NK2590 mig-17::mNG::3xFlag; shc19 Ji et al. 2019 FDU1506 Oligonucleotides Table S2 Recombinant DNA pDD122 (Peft-3::Cas9 + ttTi5605 sgRNA) Addgene 47550 mNG-C1^SEC^linker This study mNG-SEC-LL mNG-C1^SEC^bothlink This study mNG-SEC-bothlink mEos2-C1^SEC This study mEos2-SEC L4440 RNAi empty vector control Addgene 1654 mig-6 RNAi This study / Ahringer mig-6 RNAi Software and Algorithms Fiji Schindelin et al., 2012 Image J 2.0.0-rc-69/1.52p, http://fiji.sc/ Photoshop Adobe Version CC 2018 Imaris Bitplane Version 9 Illustrator Adobe Version CC 2018 R GNU Project Open Source μManager Edelstein et al., 2010 Version 1.4, https://micromanager.org/ iLas Biovision V.1.2.3 MetaMorph Molecular Devices 7.10.3.279 Excel Microsoft Version 15.39 GraphPad Prism GraphPad Version 7 Open in a separate window KEY RESOURCES TABLE In vivo visualization of 29 basement membrane (BM) proteins fused with mNeonGreen Embryonic BMs share a common template but diversify greatly during development Stable BM proteins form a scaffold that supports movement of dynamic BM components Papilin promotes local BM collagen removal by limiting GON-1 protease localization

Techniques: Virus, Recombinant, Mutagenesis, CRISPR, Knock-In, Clinical Proteomics, Membrane, Plasmid Preparation, Control, Software

Journal: Developmental cell

Article Title: Comprehensive endogenous tagging of basement membrane components reveals dynamic movement within the matrix scaffolding

doi: 10.1016/j.devcel.2020.05.022

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: L4440 RNAi empty vector control , Addgene , 1654.

Techniques: Virus, Recombinant, Mutagenesis, CRISPR, Knock-In, Clinical Proteomics, Membrane, Plasmid Preparation, Control, Software