...editing window specified by the gRNA. Adenine base editors convert adenine to inosine, which is replaced by...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance...preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism which frequently causes insertions... repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted...your gene of interest’s promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re ... based on factors such as selectable marker or cloning method. When using CRISPR, you will need to express... same cell at the same time. Single plasmids containing both the gRNA and Cas protein act as all-in-one...

...editing window specified by the gRNA. Adenine base editors convert adenine to inosine, which is replaced by...with a wide variety of Cas-containing plasmids. ID Plasmid Promoter Cloning Enzyme Co-expressed Cas9 Cas9...preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism which frequently causes insertions... repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted...your gene of interest’s promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re ... based on factors such as selectable marker or cloning method. When using CRISPR, you will need to express... same cell at the same time. Single plasmids containing both the gRNA and Cas protein act as all-in-one...

... in one cloning step rather than two. Stephen Ekker Golden Gate TALEN 2.0 Golden Gate cloning method. ...Keith Joung Zhang Lab TALE Toolbox PCR/Golden Gate cloning method. Optimized for human expression. Feng Zhang... Assembly Kit Assembly by ligation-independent cloning (LIC). Validated in human cells. Veit Hornung Musunuru...RR) used are obligate heterodimers that require cloning of left and right TALEN monomer proteins into opposite...are mammalian expression/mRNA synthesis vectors containing unique shorter N- and C-terminal domains (N153AA...fluorescent TAL effectors to visualize subcellular positioning of target DNA sequences in living cells (TALE-mediated...TALEN kit, (ii) a lacZ fragment for blue/white-screening in E.coli , (iii) CAG promoter and Kozak sequence...

... expression, Ligation Independent Cloning (LIC) Bacteria Lac, T7, araBAD p15TV-L - His-tagged... Plasmids - More LIC bacterial cloning vectors pTD plasmid ... pACU2 - Modified pUAST vector containing both P-elements and attB site for ...GFP-tag fusions for transcription pCSDest - Easy cloning of your gene using Gateway recombination... protein. Just remember that when you are designing your plasmid you should keep your gene "in... GFP for mammalian expression pcDNA3 GFP LIC cloning vector - C-terminal GFP for mammalian... expression Puromycin Mammalian pLKO.1 - TRC cloning vector - Lentiviral shRNA expression...

...- Barbier Lab Cloning and Expression Bacteria and yeast expression vectors : FX cloning System - Geertsma...Parts Kit : Molecular cloning for E. coli - Densmore Lab EcoFlex MoClo : Modular cloning for applications ...Microbiology : Sets of plasmids for fluorescence, cloning and expression, and synthetic biology. CRISPR Resources...sets of plasmids intended for use together (e.g. cloning and expression systems) and therefore supplied ....0 Yeast Toolkit - Borodina Lab Golden GATEway cloning system - Wittbrodt Lab MultiSite Gateway Kit : ...predicted open reading frames for vaccinia virus - Bennink and Yewdell Labs Yarrowia lipolytica Golden Gate...

...to add the CACC and AAAC cloning overhang. When designing oligos for cloning my target sequence into a...published with thorough step-wise instructions on designing and using Cas9-based genome engineering tools ...Should I add the PAM sequence to the oligo when cloning my target spacers into the PX330 vector backbone...guidelines? A few notes below are considerations for designing HR donor. Generally if off-target (non-specific... the Bench FAQs How can I quickly check if the cloning of my oligo into a backbone like PX330 worked? ...Why does the PX260 plasmid use 30nt oligos for cloning but PX330 only uses 20nt? The reason for the 30nt...the construct is from our 'Chimeric RNA' design (meaning the mature processed form of crRNA is fused chimerically...

...editing window specified by the gRNA. Adenine base editors convert adenine to inosine, which is replaced by...preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism which frequently causes insertions... repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted...your gene of interest’s promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re ... based on factors such as selectable marker or cloning method. To isolate mammalian vectors, simply type... same cell at the same time. Single plasmids containing both the gRNA and Cas protein act as all-in-one...protein can be paired with a wide variety of Cas-containing plasmids. Do you have suggestions for other plasmids...

..., Terns MP, White MF, Yakunin AF, Garrett RA, van der Oost J, Backofen R, Koonin EV. 2015. An updated ...The CRISPR adaptive immune system seems like a winning strategy to ensure prokaryotic survival, but it...Cas9 could be repaired through non-homologous end joining (NHEJ) , an error-prone process that produces insertions...sequencing, CRISPR is a robust system for genome-wide screening. CRISPR Technological Advances CRISPR technology...Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman...Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. 2015. In vivo genome editing...Zhang F. 2014. Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells. Science . 343(6166):84-7. PMID...

...useful tool for genetic screening experiments, due to the relative ease of designing gRNAs and the ability...plasmids, each containing multiple gRNAs for each target gene. In a CRISPR screening experiment, target...are then screened for a phenotype of interest. Screening experiments using a pooled CRISPR library are ...libraries and keep the following in mind: CRISPR screening experiments require electroporation to amplify...NGS) to confirm library representation. CRISPR screening experiments require large quantities of cells ...lab. Once you have determined that CRISPR-based screening is appropriate for your experiment, the next step...Streptococcus pneumoniae 170432 Knockout S. pneumoniae Veening N/A 1 1,499 MusCK Library 174196 (Library A) 174197...

...Additional Cloning Information Cloning Protocols Single guide cloning: Zhang Lab General Cloning Protocol...provided as empty backbones that utilize Golden Gate cloning to insert the desired guide RNA sequence by digestion... annealed and phosphorylated oligonucleotides containing the guide target sequence: Up to two guides can...Protocol (PDF, 237 KB) Dual guide cloning: Brzezinski Lab Dual Guide Protocol (PDF, 990 KB) Plasmid and Sequence...

...Proteins FREQ-Seq Kit FX Cloning Kit GreenGate Cloning System (Plant) MoClo Modular Cloning Modular PUF Assembly...Plasmids by Category Cloning and Genomic Tools Browse plasmids related to cloning and genomic modification...Pamela Silver Lei Stanley Qi Jeff Tabor Jan-Willem Veening Christopher Voigt Clifford Wang Ron Weiss Synthetic... Networks and Gene Regulation Browse plasmids containing both naturally occurring and synthetic regulatory...

...Packaging Service page . Pooled CRISPR Libraries for Screening Use pooled CRISPR libraries to screen for genes...backbone lentiGuide-Puro targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000 non-targeting...backbone lentiCRISPRv2 targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000 non-targeting... XPR_502 (P65 HSF) targeting 18,885 genes and containing 56,762 unique sgRNAs. This backbone does not ...backbone pXPR_050 inhibiting 18,901 genes and containing 57,050 unique sgRNAs along with 500 non-targeting...backbone pXPR_050 inhibiting 18,899 genes and containing 57,011 unique sgRNAs along with 500 non-targeting...Mouse sgRNA library in backbone lentiGuide-Puro containing 78,637 unique sgRNAs targeting 19,674 genes along...

...electrophoresis (PAGE) followed by silver staining or SYPRO Ruby staining and the molecular weight and relative...after negative staining. Empty vector particles can be identified after negative staining and appear darker...purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV subjected to ...Microscopy The ratio of empty to full (i.e., genome containing) AAV particles within representative vector preparations...Trono lab . Cell culture medium (OptiPro SFM) containing lentivirus is first cleared by low-speed centrifugation...

...Components for a tetracycline off system are: Plasmid containing a tetracycline-dependent promoter upstream of...Components for a tetracycline on system are: Plasmid containing a tetracycline-dependent promoter upstream of...tetracycline or its derivatives which can result in unintended system induction. Investigate whether or not ...Lentiviral vector for inducible expression for Gateway cloning; selection cassette in format: PGK-rtTA-2A-puro...shRNA; see article for detailed information about cloning and additional plasmids tetR-KRAB On Aebischer ...Elledge 35625 pAAV-Ptet-RFP-shR-rtTA AAV; shRNA cloning vector; for evaluation of shRNA efficacy using ...Sleeping Beauty transposon system; has luciferase in cloning site; see article for additional selection and ...

...soon containing SARS-CoV-2 sequences. SARS-CoV Plasmids : Plasmids that are available containing SARS-...called SARS-CoV-2. Scientists have been working at lightning speed to elucidate as much as they can about this...classic) sequences. Mammalian Targets : Plasmids containing mammalian sequences linked to SARS-CoV-2 infection...). Plasmids SARS-CoV-2 Plasmids Many plasmids containing SARS-CoV-2 sequences are now available and more...-2 Spike Receptor Binding Domain Deep Mutation Scanning Library - Library of Spike protein RBD mutations...Access Collections of COVID-19 related articles spanning multiple publishers can be found here: ASM COVID...

...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance...preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism which frequently causes insertions... repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted... based on factors such as selectable marker or cloning method. When using CRISPR, you will need to express... same cell at the same time. Single plasmids containing both the gRNA and Cas protein act as all-in-one...

...506 517 93 5.7 10 min Monomer pCS2+mNeonGreen-C Cloning Vector - Mammalian Expression Jump to Top Yellow...529 59 5.7 Monomer (A206K) pcDNA3 mCitrine LIC cloning vector (6C) - Mammalian Expression pCS2+8CmCitrine.../Xenopus/C.elegans/Sea urchin pET mCitrine LIC cloning vector (u-mCitrine) - Bacterial Expression mCitrine-N1... 548 562 49 6.5 2.5 hr Monomer pET mOrange LIC cloning vector (u-mOrange) - Bacterial Expression pCS2+...pmScarlet_C1 - Mammalian Expression pCS2+mScarlet-C Cloning Vector - Mammalian Expression mScarlet-I 569 593...587 610 16 4.5 15 min Monomer pcDNA3 mCherry LIC cloning vector (6B) - Mammalian Expression pLV-mCherry ...Zebrafish/Xenopus/C.elegans/Sea urchin pET mCherry LIC cloning vector (u-mCherry) - Bacterial Expression pFA6a-link-yomCherry-SpHis5...

...well as by our trusted partner labs. Learn More Cloning and Expressing Antibodies Antibody Plasmids Antibodies...synthetically produced recombinant antibodies, created by cloning antibody components into plasmids and expressing...and their applications in our Antibody Guide ! Cloning and Expressing Antibodies The following table highlights...mammalian expression vector A tool kit for rapid cloning and expression of recombinant antibodies. Sci Rep... Rep. 2014 Jul 30;4:5885. Andrew Beavil Cloning and mammalian expression of mouse and human IgG monoclonals...and secreted proteins suitable for wholemount immunostaining. Biochem Biophys Res Commun. 2015 Jan 2;456...

...targets adenenine, NG targets thymine, and NN targets guanine (though NN can also bind adenenine with lower...approximately every 35bp. Researchers are still determining the importance of context for each TAL effector...be used with the recently developed Golden Gate cloning method to assemble “multiple DNA fragments in an...to hone these tools and the researchers who are learning to wield them. Learn more about Addgene’s TAL ...

... when cloning in E. coli . This alteration enables the generation of a single vector containing Cre and...Blog: Cre-lox Addgene's Blog: FLEx Vectors Cre-containing Plasmids loxP Constructs Cre Reporters & Tools...Cre alternatives from Gerald Rubin's lab. Cre-containing Plasmids Scientists have developed ways to tightly...deleting the floxed portion of a construct during the cloning process). VCre and SCre are alternative Cre recombinases...pAAV.cTNT.iCre iCre and tdtomato Chicken cardiac troponin T AAV Pu 70120 pEMS2159 iCre with MCS for inserting...132551 AAV-U6-gRNA-TnT-Cre Cre Chicken cardiac troponin T AAV Pu 134308 pCAG-Synaptophysin-TdTomato-IRES2...the expression of one gene while simultaneously turning on another. The FLEx switch system takes advantage...