... Plasmid Collections Plant Plasmids Plant Plasmids and Resources...creating plant expression vectors: A modular plasmid assembly kit for multigene expression, gene silencing...full selection of plasmids expressing different functional variants of Cas9 and empty gRNA backbones. A few...addition to individual plasmids, Addgene carries several useful toolkits and plasmid collections for use ...includes plasmids containing promoters that drive general and species-specific gene expression (see the... - deposit plant plasmids with Addgene! CRISPR Please refer to Addgene's CRISPR Guide for a general introduction...highlights plasmids that can be used as empty backbones for your gene of interest for expression in plants...

...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag... for CRISPR plasmids for transcriptional activation. Plasmids are available for expression in mammalian...If the plasmid that you choose does not also express a gRNA, you will need to use a separate gRNA expression...CRISPR plasmids for activating or upregulating gene expression in various model organisms. CRISPR...Activate CRISPR Plasmids: Activate Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Publication Drosophila ID Plasmid Gene/Insert Promoter PI Publication Plant ID Plasmid Gene/Insert Promoter...

...create a single gRNA-expressing plasmid, but it can also be adapted to express multiple gRNAs. Addgene has...  Check out more Plasmids 101 Posts! Check out Addgene’s restriction cloning guide! CRISPR 101: Multiplex...oligonucleotides specifying a gRNA target sequence into a Cas9-containing plasmid such as pX330. This cloning...Multiplex Expression of gRNAs Choosing a Molecular Cloning Technique  ...Addgene’s plasmids are used with a wide variety of restriction enzyme-based cloning methods. Each method...Entry DNA overhangs may be present in the original plasmid (Option 1) or added using PCR-based amplification...fragments to create TAL effector nucleases. These plasmids were designed to utilize the BsaI and BsmBI Type...

... single protein is expressed, with the intent of purification, it will be expressed along with all other...Addgene’s antibody guide Antibody protocols  Resources on the Addgene blog Plasmids 101: Protein tags...other cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually...about that protein that no other protein in the expression system has. This is where affinity tags come ...peptides which do not naturally exist in most expression systems that can be added to the N or C terminus...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox and...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...changes. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression...CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have..., nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...(DSBs) at a specific location based on a co-expressed gRNA-defined target sequence. DSBs are preferentially...

...and lentiviral expression vectors (Table 1). The four systems Name Function Plasmid Transduction...Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a gRNA. These... Your gRNA for CRISPR Genome Editing CRISPR Cheat Sheat Resources from Addgene.org CRISPR Guide CRISPR...  The X-CHIME deposit also includes pXPR_071, a plasmid that leads to higher titer lentivirus preparations...first generation CHIME (pXPR_053). Find X-CHIME plasmids here!   Marty LaFleur is a Postdoctoral Fellow...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have...than NHEJ. ID Plasmid Gene/Insert Vector Type Promoter PI Publication Empty gRNA Expression Vectors Select...Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When using..., nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with...transcription none, need Cas9 plasmid Joung Do you have suggestions for other plasmids that should be added to...

...and Myers Tagging Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor... Tagging Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...reaction produces a single plasmid targeting vector that encodes both a guide RNA transcript and a repair...the remaining 5 plasmids include Cre and FLP expression vectors, a general cloning plasmid, and a prebuilt...PX458 . Multiple gRNA plasmids may be used. The HDR (homology directed repair) donor plasmid with homology..., which expresses an untagged Cas9 AND a gRNA targeting the AAVS1 locus. Doyon Tagging Plasmid: 3xFLAG...

...Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such ...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...variety of Cas-containing plasmids. ID Plasmid Promoter Cloning Enzyme Co-expressed Cas9 Cas9 System Selection...base-to-base conversions using a prime editing guide RNA (pegRNA). The pegRNA directs the nickase to the target site...If the plasmid that you choose does not also express a gRNA, you will need to use a separate gRNA expression...If the plasmid you’re using does not also express a gRNA, you will need to use a separate gRNA expression...CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have...

...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...CRISPR expression plasmids that have been designed for use in parasites. CRISPR...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Eukaryotic... CRISPR genome-wide knockout gRNA pooled library designed with 10 guides against 8,158 predicted protein-coding...Plasmids: Parasites Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit Transcriptional...development. This collection brings together some of the plasmids that have been used in these efforts. The content...

...Cas proteins then express and process the CRISPR loci to generate CRISPR RNAs (crRNAs). Through sequence...guanosine. RNA editors : Cas13 fused to the ADAR2 deaminase convert adenosine to inosine on RNA. Inosine...check out Addgene’s CRISPR Guide and CRISPR blog posts , or find CRISPR plasmids for your research. Addgene...CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol . 32(3):279-84. PMID: 24463574...Weissman JS, Qi LS. 2013. CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes. Cell... JA, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity... M, DiCarlo JE, Norville JE, Church GM. 2013. RNA-guided human genome engineering via Cas9. Science . ...

...As a plasmid repository, we are no strangers to cloning. If you’re reading this, then chances are you... multiple parts transcribed together. They can express one or more gene(s), and typically begin with a...for more complex CRISPR experiments (e.g., many guides/parts delivered) in certain model organisms. In...Resources on Addgene.org Addgene’s Modular Cloning Guide EcoFlex MoClo Kit CIDAR MoClo Kit MoClo Plant Parts...NEB’s DAD Interview with Nicola Patron on MoClo Plasmids 101: Golden Gate Cloning ...many gRNAs delivered with a relevant Cas nuclease. MoClo can be used to assemble multiplexed gRNA systems...PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765.  Hahn, F., Korolev, A., Sanjurjo Loures...

...structure/gene expression. RNA Targeting RNA Targeting Type VI CRISPR systems target RNA, not DNA, across...select CRISPR lentiviral plasmids. CRISPR Plasmid Kits : Find groups of plasmids that are used together ...resources or browse plasmids below by function. Addgene's CRISPR collections feature plasmids for a variety ...multiple systems. RNA Editing Cas13 enzymes fused to deaminases can edit bases in RNA. Other Applications...experiment based on a variety of factors. gRNAs gRNA plasmids to target specific genes....CRISPR plasmids available at Addgene. Search by species and CRISPR function. CRISPR... CRISPR Plasmids and Resources...

...Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression plasmids - Keith Joung Lab CRISPR/Cas9... Plasmid Collections Zebrafish Plasmids Zebrafish Plasmids and Resources... of the plasmids and tools available at Addgene for use in zebrafish. Browse Plasmids Plasmid Tools for...biosensors, and Cre expression plasmids. Large collections or kits are described below . ID Plasmid Description...resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of useful...fusion expression in organisms or cells in which an exogenous mRNA can be injected and expressed. EXPRESs...for plasmids containing your zebrafish gene of interest? Search Addgene's collection for plasmids that...

...allow the complementary base pairing between the guide RNA and the target DNA strand, and then activate two...Addgene.org Visit Our CRISPR Guide Page CRISPR/Cas9 History Browse All CRISPR Plasmids and Resources ... BioMolecular Modeling (CBM) in 1999 with the expressed purpose of creating physical models of proteins...interactive images of their proteins. Since the 1980’s, journal articles have been filled with impressive, full-color...their attention on something tangible. Under the guidance of a trained educator, these instances of focused...

...CRISPR RNA Targeting CRISPR Plasmids: RNA Targeting Browse CRISPR Plasmids By Function Genome Editing Cut ...tables below for CRISPR RNA targeting plasmids. Plasmids are available for expression in mammalian systems...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...CRISPR plasmids for targeting RNA sequences, which include the enzymes C2c2/Cas13a and Cas13b. CRISPR... target RNA rather than DNA. In bacteria, once they have recognized and cleaved the target RNA sequence...to attentuate RNA levels. In mammalian systems, Cas13a does not exhibit the collateral RNA degradation ...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacterial ID Plasmid Gene/Insert...

... (2013). Plasmid is low copy number. 45946 pU6-BbsI-chiRNA : Plasmid for expression of chiRNA under the...Lab CRISPR Plasmids flyCRISPR Plasmids Available from Addgene You may also like... CRISPR Guide CRISPR Protocols.... These plasmids are described in: Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease...window) Article Individual plasmids can be ordered via the links below: ID Plasmid 46294 pBS-Hsp70-Cas9 : ...Browse CRISPR Cas9 gRNA plasmids for drosophila research from the O'Connor-Giles, Wildonger, and Harrison...generated within a month. phsp70-Cas9 and a single chiRNA plasmid can be coinjected to generate mutations via...DSBs. Coinjection of phsp70-Cas9 and plasmids encoding two chiRNAs can induce defined deletions between...

...Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...If the plasmid that you choose does not also express a gRNA, you will need to use a separate gRNA expression...CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have..., nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with...Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid Calarco...

...fluorescent protein timers When FP timers are first expressed, they predominantly fluoresce one color but slowly...timers (whether as part of protein fusions or expressed independently) originally fluoresce green or blue...displays predictable kinetics over 14 hours of expression in C. elegans embryos. However, this initial ...ideal FP timers: 1. Ratiometric determination of expression time The ratio of mature to immature fluorescence...fluorescence from the FP timer is dependent on the total expression time but independent of protein concentration...Calibration curves correlating this ratio to the total expression time can therefore be used to determine how long... long a pool of FP timers has been expressed, either in whole cells or as a pool of fusion proteins localized...

...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...below for CRISPR plasmids to help purify genomic loci. Plasmids are available for expression in mammalian ...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...Zebrafish Xenopus CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Identifying...Catalytically inactive CRISPR plasmids for purification of target loci. CRISPR...Purify Genomic Loci Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit Transcriptional...