...with a single insertion because it will segregate 3:1 in the T2 generation. You also have no control over... transgenic plants in the process.     References 1. Emami, S., Yee, M. and Dinneny, J. R. (2013) ‘A robust...transformation’, Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829..., a graduate student at Stanford University. Arabidopsis is a fantastic model organism for many reasons... are many motivations to generate transgenic Arabidopsis, from studying transcriptional and translational... plants, to complementing mutant phenotypes. Arabidopsis is amenable to the floral drip or dip transformation...transformation protocol, as well as protocols for growing Arabidopsis are also available (Weigel and Glazebrook, 2002...

...FokI S. pyogenes Conklin pcrRNA.ind 61284 Bacteria araB none S. pyogenes Beisel pcrRNA.con 61285 Bacteria...Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO.1-puro U6 sgRNA BfuAI stuffer 50920 Mammalian/Lentiviral...Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm EcoRI none S. pyogenes de Bono...Zebrafish BseRI none S. pyogenes Zon pCRISPomyces-1 61736 Bacteria BbsI yes, cut S. pyogenes Apramycin...see plasmid page yes, cut S. pyogenes BFP Kuhn pLKO.1-puro U6 sgRNA BfuAI large stuffer 52628 Mammalian/...Puro Zoldos pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) 71707 Mammalian hU6 yes, cut S. pyogenes Draetta...expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent promoters. sgRNAs...

... expression F- ompT lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General protein expression  BLR ...folding, and deal with sub-optimal codon usage (Table 1) A few mutations are common to all or most expression...curated list of Bacterial Expression Systems Table 1: E. coli expression strains  Note: All strains are...recommended for toxic proteins BL21-A1 Tetracycline Arabinose-inducible expression of T7 RNAP; IPTG may still...expression strain to make sure they are compatible. An arabinose-inducible plasmid will not express in an IPTG ...

...AdEasier®-1 cells (strain) - Bacterial strain that contains AdEasy®-1 plasmid... expression Puromycin Mammalian pLKO.1 - TRC cloning vector - Lentiviral... Tet-inducible lentiviral shRNA expression pMKO.1 puro - Retroviral shRNA expression ... expression Hygromycin Mammalian pLKO.1 hygro - Lentiviral shRNA expression... - Retroviral gene expression pMKO.1 GFP - Retroviral shRNA expression pSico... expression, multiple selectable markers pMKO.1 puro - Mammalian retroviral shRNA expression...under your chosen promoter pAdEasy®-1 - Recombine plasmids from the shuttle...

...Cre-GFP fusion EF-1 alpha Mammalian Sauer 11923 pBS598 EF1alpha-EGFPcre Cre-EGFP fusion EF-1 alpha Mammalian...mCherry coexpression EF-1 alpha AAV Deisseroth 55635 pAAV-EF1a-sCre SCre EF-1 alpha AAV Deisseroth 55636...55636 pAAV-EF1a-Cre Cre EF-1 alpha AAV Deisseroth 55638 pAAV-EF1a-vCre VCre EF-1 alpha AAV Deisseroth 59701...) EF-1 alpha AAV Cepko 69571 pAAV-EF1a-C-CreintG Cre recombinase dependent on GFP (CRE-DOG) EF-1 alpha... Mammalian Sauer 11918 pBS513 EF1alpha-cre Cre EF-1 alpha Mammalian Sauer 11919 pBS448 RSV-GFPcre Cre-...11955 pBS505 EF1alpha-EGFPcre* Cre-EGFP fusion EF-1 alpha Mammalian Sauer 11956 pBS594 promoterless EGFPcre...GFAP-Cre Cre GFAP Mammalian Sofroniew 25997 LV-Cre pLKO.1 Cre and shRNA coexpression CMV Lentiviral Fuchs 26646...

...PDF 2.3 MB Zhang GeCKO pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9...off-target binding. Currently supports: Drosophila, Arabidopsis, zebrafish, C. elegans, mouse, human, rat, cow...inactivation, NHEJ, and HDR. Reference genomes include Arabidopsis, C. elegans , sea squirt, cavefish, Chinese hamster...zebrafish, fly, worm, Japanese rice fish, maize, Arabidopsis, and rice. Proof-of-concept performed in zebrafish...zebrafish, sea squirt, Drosophila, C. elegans, Arabidopsis, rice, sorghum, silkworm, and budding and fission...off-target binding. Currently supports: Drosophila, Arabidopsis, zebrafish, C. elegans, mouse, human, rat, rabbit...off-target binding. Currently supports: Drosophila, Arabidopsis, zebrafish, C. elegans , mouse, human, rat, yeast...

...expression inducible by red light. pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline...Wolf Frommer Reporter for arabinose levels in bacteria. The presence of arabinose enhances the FRET signal...14030 FRET Patrick Daugherty FRET: Vector for arabinose inducible expression of an N-terminally His-tagged...14031 FRET Patrick Daugherty FRET: Vector for arabinose inducible expression of an N-terminally His-tagged... LIC Series (8-Series) Various Plasmids pBAD (arabinose inducible, glucose repressible) His, Strep, mOCR...TEV cleavage sites to remove tags Scott Gradia Arabinose inducible vectors that come with various epitope...Various Plasmids pBAD, OR2-OR1-PR, pLtetO, pLlacO Arabinose, Anhydrotetracycline, lactose, IPTG Richard Murray...

...Hyg Chen 86196 pYPQ141-ZmUbi-RZ-As Maize ubiquitin 1 none As Cpf1 Qi 91715 pKEE401 yes, cut S. pyogenes...U6 BbsI none S. pyogenes Gao 59188 pBlu/gRNA Arabidopsis U6 BbsI none S. pyogenes Stupar 62200 pBUE411...

...Element On or off PI 26429 pLenti CMV rtTA3 Blast (w756-1) Lentiviral reverse tetracycline-controlled transactivator...toxicity, a known half-life (24 hours), and is compararably inexpensive. The induction profiles for other...

...a wide range of inducer concentrations (0.1-1% L-arabinose) without signs of toxicity or burden relative...bacteria have colonized (Fig. 1c).     Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram...ultrasound contrast. In all the cell lines we tested, 1 µg/mL doxycycline was sufficient to saturate expression...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the ... tumors and tumor-homing bacteria. Nat Biotechnol 1–13 (2023) doi:10.1038/s41587-022-01581-y  ...for longer. Cultures induced with arabinose and colonies on arabinose-containing plates will be visibly... inducible expression systems in E. coli. The arabinose-inducible promoter pBAD and a low-copy number ...

...in all other plants, anthocyanins.   Figure 1: An adult plant expressing RUBY. Photos by Andrew ... be collected from heterozygous individuals. Only 1/4 of the descendants of the heterozygous plants, therefore... (2020). https://doi.org/10.1038/s41438-020-00390-1 Additional resources on the Addgene blog  CRISPR 101...occurs in T0 plants (from callus) or T1 plants (Arabidopsis floral dip). In the next generation the CRISPR...planning to use it to simplify my work with an Arabidopsis mutant that can’t set seeds. I study the development...   Fig. 3: (Top right) A RUBY-expressing Arabidopsis seed following floral dip transformation next...without antibiotics. (Top left) Mature silique (Arabidopsis fruit) with fertilized developing seeds from ...

...Table 1 below outlines a few of the more common genetic changes found in E. coli strains. Table 1: Common...amber (UAG) stop codon by tyrosine insertion λ-thi-1 or thi1 Mutation in thiamine metabolism Requires exogenous... all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain Natural...plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF...) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109   General cloning and...strain for cloning repetitive DNA. endA1 glnV44 thi-1 relA1 gyrA96 recA1 mcrB+ Δ(lac-proAB) e14- [F' traD36...of an E. coli K-12 strain. F- λ- ilvG- rfb-50 rph-1 NEB Stable   For cloning into and storage of lentiviral...

...oxygen. Additionally, its status as a Biosafety Level 1 microbe makes it a perfect candidate for introductory...Transport, biofilms, and much more. Because of it’s BSL-1 status, E. coli can be used as a model to relate these...with superior performance. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-04899-x Yokoyama...is as a biomedical model organism.  Arabidopsis thaliana Arabidopsis thaliana is one of, if not the most...community.  The Arabidopsis genome project marked the beginning of widespread Arabidopsis genetics research...characterized genome, and widespread utilization, Arabidopsis has made significant contributions to understanding...understanding plant and crop science alike.  Arabidopsis has many of the traits that make an ideal model organism...

... that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain...expression needed for most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize...AraC system, which uses arabinose, requires many wash steps to remove arabinose making it difficult to ...🎶 “Bye bye L-arabinose drivePut your cultures in the shakerTurn the LEDs onAnd when you wantYou can ...recently published in @nchembio Title: Bye bye L-arabinose drive. Dedicated to Edo, Armin and @KhammashLab...lab,” says Di Ventura. It relies on the sugar arabinose to activate transcription by causing conformation...to regulate quickly. Arabinose also circulates within growth media and can’t be contained to a specific...

...where she focuses on (1) salt-induced changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns...the journal-specific guidelines.  Peer-review step 1: The first read At the first read of the paper, I ... of Arabidopsis HapMap population in Amsterdam. Currently, she is a postdoc at KAUST, Saudi Arabia, in...Julkowska, a postdoctoral researcher at KAUST, Saudi Arabia. From the perspective of an author submitting ... specific repositories (Addgene, WormBase, Arabidopsis.info). These repositories will ensure that the ...guest blogger Magdalena Julkowska from KAUST, Saudi Arabia. Magdalena Julkowska did her Ph.D. on salt-stress...

...General Design Figure 1: Two proteins are fused to the FP(11) and FP(1-10) fragments. When Protein...and 11 to yield large (1-10) and small (11) fragments. In several cases an FP(1-10) fragment may be compatible...Description Article PI Blue EBFP2 EBFP2(1-10) and Capri(1-10) for use with GFP(11) Multiplexed labeling...Commun. 2017 Bo Huang mNeonGreen3 mNG3K(1-10) and mNG3A(1-10) for use with mNG2(11) Improved yellow-...sfCherry2 sfCherry2(1-10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1-10) Improved split...brings the pieces of the FP back together (Figure 1). This method is generally known as Bimolecular Fluorescence...even from a different base FP (for example, EBFP2(1-10) can be used with GFP(11)). Some of the articles...

...dsRNA via an RNase III enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of...in the biogenesis of different small RNAs (Figure 1). These siRNAs can exert their gene silencing effect...modifications of complementary gene sequences (Figure 1). As a result, gene silencing can be easily induced...the ease of use in dsRNA applications. Figure 1: Schematic of an RNAi-inducing transgene. Typically...complementary to your gene target as inverted repeats (Figure 1). With this technique, multiple genes can be simultaneously...selection of plants undergoing RNAi, and LIIbeta F 1-2 RNAi, which allows assembly of intron-spliced hpRNAs...amiRNAs based upon miRNA precursors of rice and Arabidopsis species, respectively. Short hairpin RNA (shRNA...

...new combinations of PE protein components (Figure 1), an altered epegRNA delivered with an enhanced pU6cm...amplification of cargo, and heat treatment.  Figure 1: Left: recombined prime editing tools PE2max-NC and...conformational dynamics of the motor protein kinesin-1, the Twelvetrees Lab took advantage of the self-labeling..., Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and adopts an open conformation... to drive prime editing in both tomatoes and Arabidopsis, suggesting the improvements may be broadly applicable...enables heritable desired edits in tomato and Arabidopsis. Nature Plants, 10(10), 1502–1513. doi: https...

...Hsp70, the heat shock releases heat shock factor 1 (HSF-1), which subsequently binds to heat shock elements...used for bacterial protein purification. When arabinose is absent, regulatory protein AraC binds O and...pBad, blocking transcription. The addition of arabinose causes AraC to bind I1 and I2 sites, allowing ...allowing transcription to begin. In addition to arabinose, cAMP complexed with cAMP activator protein (CAP) can...

... medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our research is to enhance ...activities in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. Lee, Min Woo, and Yinong...   C. roseus is not a model organism such as Arabidopsis or tobacco. Studying gene function in C. roseus...method for overexpressing genes In plants like Arabidopsis and tobacco, Agrobacterium infiltration into ...expression assay by agroinfiltration of leaves." Arabidopsis Protocols. Humana Press, 2006. 225-229. PubMed...transient gene expression analysis in seedlings of Arabidopsis and other plant species." Plant methods 5.1 (...of our plant biology blog posts Find tips for Arabidopsis transformation Read our Plasmids 101 blog posts...