...Reporter Gene Guide RNA expression plasmids for endogenous human genes Guide RNA expression plasmids for endogenous... CRISPR-Cas/RGN expression plasmids for use in zebrafish Guide RNA expression plasmids for the EGFP Reporter...CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids You may also like... CRISPR Guide CRISPR Protocols gRNA Design...specificities Dimeric CRISPR RNA-guided FokI Nucleases (RFNs) CRISPR-Cas/RGN expression plasmids for use in human ...CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids Genome...consist of two components: a short ~100 nt guide RNA ( gRNA ) that uses 20 variable nts at its 5’ end ... recently described gRNA and Cas9 expression vectors and validated the use of RNAs produced by these vectors...

...with the guide RNA. For most applications, we recommend using Cas9 with the single guide RNA (sgRNA), which...SpCas9n, D10A nickase) + single guide RNA: These plasmids contain two expression cassettes, a human codon-optimized...SpCas9-2A-EGFP and single guide RNA 62988 : PX459; SpCas9-2A-Puro and single guide RNA 48873 : PX460; SpCas9n...) and single guide RNA 48140 : PX461; SpCas9n-2A-EGFP (D10A nickase) and single guide RNA 62987 : PX462...FACS sorting SaCas9 + single guide RNA: These plasmids contain two expression cassettes, a human codon-optimized...D10A nickase) + CRISPR RNA array + tracrRNA: This plasmid contains three expression cassettes. To target...both the SaCas9 gene and its single guide RNA (sgRNA) into one plasmid (Ran et al., Nature 2015). The dual...

... the RNA polymerase, thus strongly decreasing the expression level. Mismatches between the guide RNA and...range of expression levels. This collection comprises 20 plasmids, each carrying two CRISPR guides. The first... can be controlled using a plasmid‐borne CRISPR array coding for guide RNAs (diamonds) interspaced with... CRISPR Repression Plasmids You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR...From the Bikard Lab, CRISPR plasmid collection for modulating expression levels of two targets independently...arrays to modulate the expression levels of two targets independently. The RNA-programmed DNA-binding ...nucleotides. Together, these plasmids allow to explore the whole range of expression levels for these two reporters...

...iChIP plasmid, expresses 3xFNLDD in mammalian cells 48972 3xFNLDD/pMXs-IG iChIP plasmid, expresses 3xFNLDD...interest 61079 gRNA-hIRF-1 #12 Expresses a guide RNA (gRNA) to target human IRF-1 promoter 61080 gRNA-hIRF-1 ...regions of interest. 98134 hIRF-1 gRNA #409 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter...Sa-Cas9 system. 105283 hIRF-1 gRNA #351 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter...1 #12/pSIR Expresses a guide RNA (gRNA) to target human IRF-1 promoter 62190 3xFLAG-dCas9/pTEF1p-CYC1t...scheme Fujii Lab Plasmids Individual plasmids can be ordered via the links below: ID Plasmid Description 47948...pMD20 iChIP plasmid, LexA-binding elements 48874 3xFNLDD/pCMV-7.1 iChIP plasmid, expresses 3xFNLDD in ...

...Science Guides CRISPR Guide CRISPR Guide Jump To: CRISPR Guide Overview CRISPR Basics Small Edits Large...that target RNA instead of DNA enables tracking of RNA in live cells, including orthogonal RNA/DNA labeling...) and editing (B) with Cas13 Browse Plasmids: RNA Targeting , RNA Editing Degrading DNA with Cas3 Rather...Repurposing CRISPR as an RNA-Guided platform for Sequence-Specific control of gene expression. Cell . 152 (5),...components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme). The gRNA is a short...conversions using a prime editing guide RNA ( pegRNA ). Like sgRNA, pegRNA directs the nickase to the target...delivering multiple gRNAs using a single plasmid ensures that all gRNAs will be expressed in the same cell...

...Activator Expression Plasmids ID Plasmid 48223 pAC91-pmax-dCas9VP64 : dCas9VP64 on pmax expression vector...containing the sgRNA expressing fragment produced by PCR), or expressed from a single dual-expression vector ... Plasmid 48236 pAC2-dual-dCas9VP48-sgExpression : Dual expression construct expressing both dCas9VP48 ...single guide RNA (sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component...dual-dCas9VP160-sgExpression : Dual expression construct expressing both dCas9VP64 and sgRNA from separate...CRISPR Plasmids Available from Addgene You may also like... CRISPR Guide CRISPR Protocols gRNA Design ...Individual plasmids can be ordered via the links below: Table 1. dCas9 Activators sgRNA Dual Expression ID Plasmid...

...BFP fusion expression plasmid 44248 pgRNA-humanized A customizable gRNA expression plasmid for use with...bacterial Cas9 expression plasmid 44250 pwtCas9-bacteria A wild-type Cas9 expression plasmid for use editing... epeats) pathway, as an RNA-guided DNA binding platform, to repress expression of arbitrary genes in bacteria...fluorescent genes. These plasmids are described in: Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific...resistance. The second plasmid ( pgRNA_bacteria ) contains a constitutive sgRNA expression cassette on a ColE1...bacterial genomes 44251 pgRNA-bacteria A customizable gRNA expression plasmid for use with the bacterial... CRISPR Qi Lab CRISPR Plasmids Qi Labs CRISPR Plasmids Available...

...find plasmids that can be used to engineer various types of viruses. Lentivirus Plasmids | Guide Lentiviruses... high but transient transgene expression. γ-Retrovirus Plasmids | Guide γ-retroviruses, commonly referred...enabling long term transgene expression. Overview of Viral Plasmids While plasmids are typically used to encode...Viral Plasmids and Resources You May Also Like... Addgene's Viral Service Biosafety Guide Blog: Plasmids...eBook Overview of Viral Plasmids Which Type of Virus Should I Use? Use this guide to learn more about the...for select plasmids are now available through Addgene, learn more here . Browse Viral Plasmids by Type Visit...host genome. Adeno-associated Virus (AAV) Plasmids | Guide Adeno-associated viruses (AAVs) are commonly used...

...System Promoter Guide RNA Type Cloning Enzyme Co-expressed Cas9 Selection PI 132777 pU6-pegRNA-GG-acceptor...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...below for CRISPR prime editing plasmids. Plasmids are available for expression in mammalian systems, bacteria...mutations using a prime editing guide RNA (pegRNA). As with a typical gRNA, the pegRNA is designed with a spacer...nicking sgRNA), cloning enzyme, selectable marker, and whether the plasmid contains Cas9. ID Plasmid Expression...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Drosophila ID Plasmid Gene/Insert...

...CRISPR Plasmids: Mammalian Expression The following CRISPR plasmids have been designed for use in mammalian...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...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... kit. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors You..., nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with...

...j.bbrc.2013.08.013. Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such ...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...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..., nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with...

...fusion shuttle plasmid for shuttling U6-guide cassettes to make dual guide expressing plasmids dCas9-KRAB-...dual guide construct. Plasmids 175573 and 175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571...containing the guide target sequence: Up to two guides can be expressed from the same plasmid by utilizing...utilizing one of the dual guide shuttle plasmids to insert a second guide within a U6-guide cassette. This process...159655 are single guide expressing double stranded break constructs. Plasmids 171098 and 175570 allow conversion..., 237 KB) Dual guide cloning: Brzezinski Lab Dual Guide Protocol (PDF, 990 KB) Plasmid and Sequence Derivative...advantage of the Zhang lab all-in-one (guide/Cas9/reporter) pX458 plasmid family. They have made several modifications...

...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...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...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...

...high expression, so adding this G could help with expression, though it is possible for the plasmid to ...with PX330 plasmid, clone in your target guides, test run them and then select the best guide to co-transfect...usually pick 3-6 guides around the region to find the most efficient guide (most guides would work, but...our 'Split RNA' design, where the pre-processing form of crRNA is expressed and a tracrRNA (the helper...sequence, and the concentration of Cas9 and guide RNA (gRNA). These considerations only matter if the PAM...helper RNA that facilitate the processing of the crRNA) is also expressed from a separate promoter. In the... may also like... Zhang Lab CRISPR plasmids General CRISPR Plasmids Dr. Le Cong, the first author of one...

...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag...below for CRISPR Transposase plasmids. Plasmids are available for expression in mammalian systems and bacteria...CRISPR Plasmids: CRISPR Transposases (CASTs) Browse CRISPR Plasmids By Function Genome Editing Cut Base...to target locations in the genome defined by a guide RNA. The transposase can then integrate donor DNA....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 CRISPR-associated...

...Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag... for CRISPR plasmids for transcriptional inhibition. Plasmids are available for expression in mammalian...If the plasmid you’re using does not also express a gRNA, you will need to use a separate gRNA expression...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast 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 Catalytically...

... 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...

...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...

...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...