...dTom-nlsdTom E2 regulatory element dTomato Control Dimidschstein 135631 pAAV-S5E2-GFP-fGFP E2 regulatory element...eYFP E2 regulatory element Activator C1V1 Dimidschstein 135634 pAAV-S5E2-ChR2-mCherry E2 regulatory element...Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory element; NHP, non-human primate. Image courtesy...DREADD Roth 135635 pAAV-S5E2-Gq-P2A-dTomato E2 regulatory element Activator DREADD Dimidschstein Biosensors...GCaMP Kim , GENIE 135632 pAAV-S5E2-GCaMP6f E2 regulatory element Calcium sensor GCaMP Dimidschstein 100854...

...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and ...edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed...Nickase mutants can also be used with a repair template to introduce specific edits via homology-directed...primer binding site and the desired edits on an RT template. Plasmid Gene/Insert Promoter PI Publication Activate... to direct the dCas9-activator to promoter or regulatory regions of your gene of interest. If the plasmid...

...Overview of TALEN Technology Imagine being able to manipulate a specific region of DNA in the genome of your...species almost as easily as correcting a typo in the latest draft of your dissertation. Well, that will probably...recognition code of the TAL effectors, which were isolated from the plant bacterial pathogen Xanthomonas....bind adenenine with lower specificity). With the template for studying this type of technology already laid...Intuitively, one would expect that assembling relatively small, repeat-laden DNA regions into a single...construction of sequence-specific TAL effectors for modulating mammalian transcription. Nat Biotechnol. 2011...

...Find CRISPR plasmids to study the regulatory networks involved in retinal development....Collection The Brzezinski lab investigates gene regulation in the context of the developing mouse retina...retina. To aide in their studies of the regulatory networks involved in retinal development the lab utilizes...plasmid to expand its usefulness for the lab's gene regulation and developmental studies. Key plasmid features...followed by a quick ligation of annealed and phosphorylated oligonucleotides containing the guide target...

...Scanziani Tools for Cell Ablation These AAV encode tools that can be used for cell ablation. ID Name Expression...Tetracycline Transactivators Affinity Purification Cell Ablation Tetracycline Transactivators and Inducible Tools... toxin receptor fused to GFP for studying cell ablation. 2 Jessell , Azim 45580 pAAV-flex-taCasp3-TEVp...designer pro-taCasp3 and TEVp for studying cell ablation. 5 Shah , Wells Don’t See What You’re Looking ...

...protein translation machinery. To expand the genetic code, 4 major changes to the standard translation machinery...Accessible Using Phosphoserine to Study Protein Phosphorylation Rinehart Lab reagents tRNA/Synthetase Pairs...acids. Expanding the Genetic Code In protein translation, an aminoacyl-tRNA synthetase (aaRS) loads its...binding protein to a DNA cleaving enzyme) or to regulate the activity of a protein so that it is responsive... for another. The orthogonal synthetase must aminoacylate only the orthogonal tRNA, and not endogenous...endogenous ones. Endogenous synthetases cannot aminoacylate the orthogonal tRNA. And the orthogonal tRNA has to...

...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and ...edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed...Nickase mutants can also be used with a repair template to introduce specific edits via homology-directed... to direct the dCas9-activator to promoter or regulatory regions of your gene of interest. If the plasmid...

...BACH2 H. sapiens AATGTAGCGATTGAGAGTGTGGG 71828 methylation S. pyogenes 26969735 Zoldoš Non-targeting H. .... sapiens GTAGGCGCGCCGCTCTCTAC 71830 methylation S. pyogenes 26969735 Zoldoš AAVS1 H. sapiens GGGGCCACTAGGGACAGGAT...CAGAACACCCCCATCGGCGA 72619 cut S. pyogenes 26493208 Guigo, Johnson Malat1 H. sapiens gRNA1: GAACCGGTGGGGCTGCGTCA; gRNA2: ...GGCAGGAGAGGCCAGTTGCG 72620 cut S. pyogenes 26493208 Guigo, Johnson Malat1 H. sapiens gRNA1: GCAACTTCCATTTTCAGTCT; gRNA2: ...GGAAGCCTCAGCTCGCCTGA 72621 cut S. pyogenes 26493208 Guigo, Johnson Malat1 H. sapiens gRNA1:GCTGGGGCTCAGTTGCGTAA; gRNA2:AGGTTTCTAAAACATGACGG...AGGTTTCTAAAACATGACGG 72622 cut S. pyogenes 26493208 Guigo, Johnson Malat1 H. sapiens gRNA1:GTTGAGATGAAGCTTCTTCA; gRNA2:TCAACCGTCCCTGCAAGGCT...TCAACCGTCCCTGCAAGGCT 72623 cut S. pyogenes 26493208 Guigo, Johnson Malat1 H. sapiens gRNA1: GAAACCTCGTGTAGCTATCA; gRNA2: ...

...and GEF Plasmids The ARF (ADP-ribosylation factor) family of regulatory GTPases have long been characterized...ARF family plasmids. ARF family members include regulatory GTPases, GEFs, and GAPs. Plasmid...26893300 ). Though best known for their roles as regulators of membrane traffic, they also play critical ... and mitochondria. The activity of GTPases is regulated both upstream (by GEFs, guanine nucleotide exchange...

...researchers also use bacteria like E. coli and their relatively well understood biology to answer many interesting...and mutations to the bacterial genome itself. Manipulating the genome directly can give you more subtle...interest in bacteria, add epitope tags to it, and, later, cleave those tags from the purified protein. Protein... combinations of constituitive promoters and translation initiation elements that can be used to drive...researcher Growth of strain USO hisB- pyrF- rpoZ- on plates lacking histidine and containing 3-AT Scot Wolfe...USO hisB- pyrF- rpoZ- , this promotes growth on plates lacking His and containing 3-AT. FLIParaF.Ec-250n...

...TAL Effector Kit 2.0. Stephen Ekker Platinum Gate TALEN Kit Platinum TALENs assembled by Golden Gate. Validated... and showed robust cleavage activity in several later studies. pCAG-T7-TALEN(Sangamo)-Destination vectors... the amount of Histone H3 lysine 4 (H3K4) di-methylation at the target locus ( Mendenhall et al., 2013...This epigenome editing was also shown to alter regulation of nearby gene expression. The vectors listed...

...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ...101 post for more information about HDR/repair template design. How to use CRISPR to tag your gene of ... (3xFLAG-2xSTREP) on endogenous genes for the isolation of native protein complexes. This vector serves...encodes both a guide RNA transcript and a repair template for an individual tagging event. The kit contains...plasmids in the following articles provide PCR templates for amplification of the tag (eg GFP, Flag, YFP...

...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ... these parasites. Moreover, CRISPR-dCas9 gene regulation has also been established for some of these parasites...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and ...edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed...

...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and ...edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed... to direct the dCas9-activator to promoter or regulatory regions of your gene of interest. If the plasmid...

...yes, methylation S. pyogenes EGFP Zoldos pdCas9-DNMT3A-PuroR 71667 Mammalian U6 yes, methylation S. pyogenes...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ...S. pyogenes Kamoun pUC119-gRNA 52255 Plant PCR template none S. pyogenes Sheen pRGEB31 51295 Plant BsaI...pdCas9-DNMT3A-PuroR_v2 74407 Mammalian U6 yes, methylation S. pyogenes Puro Zoldos pBLO1811_Cas9_noNLS_human...multiplexed plant genome editing and transcriptional regulation Plant Three types of plasmids available for use...

...Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting ...binding site (PBS) and the desired edits on a RT template. In the initial version of this tool, PE1 (depicted...increase activity, enhance binding between the template and PBS, increase processivity, and improve thermostability...The newly edited strand is then favored as the template for repair during heteroduplex resolution. The...

...set of plasmids for building homologous repair templates that incorporate a self-excising drug selection...nematodes, including C. elegans and C. briggsae. Other related miniMos plasmids can be found on the Jorgensen ...genetics, genomics and biology of C. elegans and related nematodes. WormBook - A comprehensive, open-access...original, peer-reviewed chapters covering topics related to the biology of C. elegans and other nematodes...

... epeats) pathway, as an RNA-guided DNA binding platform, to repress expression of arbitrary genes in bacteria...target loci. Thus, CRISPRi is a system that can regulate a genome instead of modifying a genome. 1. Two-plasmid...described in: Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression...PMC3664290 . CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Gilbert LA, Larson...

...vector plasmids available at Addgene and other related resources. ...transfer in clinical trials. These viruses have a relatively high transduction efficiency, though this infectivity... are typically used to encode, propagate, and manipulate genetic information, viruses are a powerful tool...These engineered viral genomes are also modified (relative to natural viruses) for safety through the removal...

...This means that the final product should be translated as a single string of amino acids that preserves... pDest vector that adds a N-terminal myristoylation signal For more information ... mammalian gene silencing (tet-regulated) L4440 - RNAi in C. elegans Transgenic... Plasmids - A system for Cre-regulated RNAi in vivo Dupuy Lab and Lehner...