...AM AAV Retrograde Tracing Gowan Tervo and Sarada Viswanathan 11:00 AM Circuit Tracing Ali Cetin 12:00 PM...this year’s workshop is to brainstorm practical strategies for using combinations of AAV-based molecular...Ben Deverman 2:00 PM Cre-DOG and Combinatorial Strategies Connie Cepko 2:45 PM Break 3:00 PM (Group Discussion...of data and protocols showing examples of AAV strategies for intersectional control. Please see our list...

...Addgene AAV transfer plasmids...or toxins should be handled at BSL-2. AAV can transduce both dividing and non-dividing cells with a low... AAV does not integrate into the host genome, transgene expression can be long-lived. The utility of AAV...effort directed toward expanding this capacity. Traditionally, AAV requires the presence of another "helper...viral genome (typically called cis -plasmid or transfer plasmid). Helper Plasmid ID Plasmid Serotype Description...

...in vitro transcription BsaI none S. pyogenes Mark pU6T7 71462 Mammalian/in vitro transcription BsaI none...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...yes, activate S. pyogenes Marraffini pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 47868 Mammalian U6...toolbox for multiplexed plant genome editing and transcriptional regulation Plant Three types of plasmids available...recombination. Qi Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs Zebrafish Plasmids...multiple functional gRNAs from a single precursor transcript Bullock pCRISPathBrick Bacteria Allows rapid ...

...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Human Bonifacino 3rd 6-25 11,108 Borodina Yeast Transporter Libraries 153101-153106 Knockout Yeast Borodina...Inhibition Mouse Weissman 3rd 5 Varies Human CRISPRi titration libraries 136478 & 136479 Inhibition Human Weissman...Knockout Human Superti-Furga 3rd 6 2,609 Human Transporter Knockout Libraries 213695 221409 221410 Knockout...Human Superti-Furga 3rd Varies Varies Human Transcription Factor Knockout Library 162275 Knockout Human...Draetta 3rd 3-4 17,032 arrays Mouse Cardiac Transcriptional Regulators CRISPR Knockout Library 138015 Knockout...

...are human (hg19 & hg38), mouse (mm10), and yeast (strain w303). CRISPR MultiTargeter Can be used to identify...and Wente Zebrafish: gRNA cloning, in vitro transcription, injection gRNA core ; Cas9 ; optimized Cas9...102.5 KB Jaenisch CRISPR plasmid summary for transcriptional activation dCas9 activators sgRNA dual expression...and BsmBI site for cloning in gRNA: pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) PDF 47.5 KB Vosshall...RNA array: Cas9 (pX260) or Cas9 D10A (pX334) ; tracrRNA: Cas9 (pX330) or Cas9 D10A (pX335) ; PDF 248.6...

...pAAV-syn-FLEX-splitTVA-EGFP-tTA helper virus for monosynaptic tracing; to be coinjected with pAAV-TREtight-mTagBFP2-B19G...pAAV-TREtight-mTagBFP2-B19G helper virus for monosynaptic tracing; to be coinjected with pAAV-syn-FLEX-splitTVA-EGFP-tTA...flip-excision switch, for AAV production or direct transfection, slow variant Kaspar Podgorski 135421-AAV1 pAAV-syn-FLEX-axon-jYCaMP1s...flip-excision switch, for AAV production or direct transfection, slow variant Kaspar Podgorski 135424-AAV1 pAAV-syn-jYCaMP1s...Synapsin1 promoter, for AAV production or direct transfection, slow variant Kaspar Podgorski 135630-AAV1 pAAV-S5E2...interneurons. For use in combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent...

...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...search the tables below for CRISPR plasmids for transcriptional activation. Plasmids are available for expression...

...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...PAM = NGG) 46759 pT7-gRNA T7 BsmBI In vitro transcription none, need Cas9 plasmid Chen and Wente 42250...42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid Joung Do you have suggestions for other...

... been engineered or identified to increase the spectral and kinetic properties as well as improve the ... a fluorescent protein to the opsin in order to track its expression and localization. Many other types...For a full description (including peak response spectra) of different types of opsins and optical switches...cellular activity. For example, search "Cdc42", "transcription", "Hahn", or other terms. ID Plasmid Description...

...in vivo transcrition controlled by the regulatory sequences of an RNA polymerase III transcribed U6 snRNA...can be efficiently generated in Drosophila and transmitted through the germline. Using an injection approach... : Generates crRNA for use in combination with tracrRNA. 51434 pHD-DsRed : vector for generating dsDNA...

...also carrying the tracrRNA sequence (not shown). (B) Relative GFP and RFP concentration given relatively... S. (2018). Tuning dCas9’s ability to block transcription enables robust, noiseless knockdown of bacterial... genes without cross‐talk. (A) Schematic of the strain expressing two reporters and dCas9 under a P-tet...

...Hockemeyer et al., Nat Biotechnol. 2011 ) TALE transcriptional activators in human cells ( Maeder et al., ...identify both dimeric TALEN and monomeric TALE transcriptional activator target sites and to simplify the ...sequences of interest for potential TALEN or TALE transcriptional activator target sites and then generates individual...Vectors for REAL, REAL-Fast and FLASH TALE Transcriptional Activator Expression Vectors for REAL, REAL-Fast...

...CRISPR knockout library targeting human transmembrane transporters. Feedback Please send your feedback to...carrier proteins (SLCs) are the largest family of transporters in the human genome with approximately 450 identified...overexpression library of human SLC proteins. Human Transporter Knockout Libraries 213695 221409 221410 Superti-Furga...

...understood but are critical for developing therapeutic strategies to prevent viral spread. As a Biosafety Level...mechanisms of infection and to determine methods of neutralizing the virus or reducing its infective potential...that can be used for measuring infectivity and neutralizing activity are listed below: pBOBI-FLuc - Lentiviral...Posts Viral Vectors 101: Pseudotyping Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV...

...mAb Collection . Once the plasmids are transiently transfected into mammalian tissue culture cells, the... is present in a sample--just to name a few. Traditional sources of antibodies for research purposes include...single domain antibodies for the conformational trapping of membrane proteins. Elife. 2018 May 24;7. ...

... mTORC1 Complex Symbol Name 4E-BP Eukaryotic translation initiation factor 4E, binding protein 1 Cyclin...Cyclin D1 Also known as CCND1 eIF-4E Eukaryotic translation initiation factor 4E GBL/mLST8 MTOR associated...Neurofibromin 1 PRAS40 Also known as AKT1S1; AKT1 substrate 1 (proline rich) PTEN Phosphatase and tensin homolog....2014.01.001. PubMed PMID: 24508508 . Targeting the translation machinery in cancer. Bhat M, Robichaud N, Hulea...

...plasmids is a bacterial stab (of a laboratory E. coli strain). Addgene continues to monitor its library and ... Integrative vector assembly of up to three transcription units in the oleaginous yeast Y. lipolytica ...Restoration of prototrophy in common S. cerevisiae lab strains - Ralser Lab Return to top Resources for... in bacterial genome editing is not always straightforward. Read our CRISPR Methods for Bacterial Genome...

... used to generate double-stranded breaks ( DSBs or cuts ) or single-stranded breaks ( nicks ). ID Name...Cas9 (dCas9) can be fused to a transactivator and used as a transcriptional activator . ID Name Insert Antibiotic...

...this serotype to deliver Cre-dependent transgenes in Cre transgenic lines. As part of our standard viral...serotype permits robust retrograde access and transgene delivery to projection neurons. Specific classes...titering by SYBR green qPCR , confirmation of the transfer plasmid with PCR, purity assessment by silver ...

...plasmids with dCas9 fused to FokI for creating double-strand breaks (DSBs)....Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...nuclease. When FokI dimerizes, it generates a double-strand break (DSB) at a specific sequence. Two unique ...