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  1. CRISPR Guide

    Type
    Collection
    ...naturally-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved...More commonly, Acrs inhibit multiple species of Cas enzymes, such as AcrIIA2 and AcrIIA4, isolated from...There are rare cases where Acr can work with multiple types, such as AcrVA3.1, which inhibits both Cas12a...Similarly, some Acr proteins are specific to a particular species of CRISPR enzymes, such as AcrIIA7–10, which... protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled Acr activity ... to protect against these foreign nucleic acids. Acr family members inhibit CRISPR by a variety of mechanisms...or inducing degradation of the Cas9 protein. Most Acr proteins are specific to a particular type of Cas...

  2. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...editing from ~30% in control samples to 0-10% in Acr samples. Acr proteins can also be used to prevent dCas9...might not be as effective. This is where Acr proteins come in. Acr proteins are small enough that they could...specific site. 2. Temporal, spatial, or conditional control of CRISPR activity Regulating Acr proteins by..., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered that...in 2012 (Bondy-Denomy et al., 2012). Anti-CRISPR (Acr) proteins are phage-derived small protein inhibitors... the CRISPR-Cas immune system of bacteria. These Acr proteins were originally found in type I CRISPR-Cas...Cas9, Cas12, and Cas13 (Marino et al., 2020). Some Acr proteins are specific to a particular Cas protein...

  3. Immunology Research Plasmids and Resources

    Type
    Collection
    ...immunoglobulin heavy diversity 2-15 D2, IGHD215 IGHD2-2 immunoglobulin heavy diversity 2-2 IGHD22 IGHD2-21 immunoglobulin...heat shock 70kDa protein 2 HSP70-2, HSP70-3 HSPA4 heat shock 70kDa protein 4 APG-2, HS24/P52, MGC131852, ...variable 2-33 (non-functional) IGLV233, V1-9 IGLV2-8 immunoglobulin lambda variable 2-8 IGLV28, V1-2 IGLV3...beta 4 DEFB-2, DEFB102, DEFB2, HBD-2, SAP1 EDN1 endothelin 1 ET1, HDLCQ7 EDN2 endothelin 2 ET2, PPET2 ...receptor subfamily 2, group C, member 1 TR2 NR2C2 nuclear receptor subfamily 2, group C, member 2 TAK1, TR2R1...suppression of tumorigenicity 2 - STC1 stanniocalcin 1 STC STC2 stanniocalcin 2 STC-2, STCRP TAC1 tachykinin,...factor 2 (granulocyte-macrophage) GMCSF, MGC131935, MGC138897 CSF2RA colony stimulating factor 2 receptor...

  4. CRISPR Guide

    Type
    Guide
    ...naturally-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved...More commonly, Acrs inhibit multiple species of Cas enzymes, such as AcrIIA2 and AcrIIA4, isolated from...There are rare cases where Acr can work with multiple types, such as AcrVA3.1, which inhibits both Cas12a...Similarly, some Acr proteins are specific to a particular species of CRISPR enzymes, such as AcrIIA7–10, which... protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled Acr activity ... to protect against these foreign nucleic acids. Acr family members inhibit CRISPR by a variety of mechanisms...or inducing degradation of the Cas9 protein. Most Acr proteins are specific to a particular type of Cas...

  5. Genetic Code Expansion

    Type
    Collection
    ...48696 pANAP AnapRS E. coli 3-(6-acetylnaphthalen-2-ylamino)-2-amino-propanoic acid (Anap) Mammalian TAG Peter...Mammalian TAG Huiwang Ai 73544 pEvol-pAcFRS.2.t1 pAcFRS.2.t1 E. coli p-acetyl-l-phenylalanine (pAcF) Bacterial...Bacterial TAG Farren Isaacs 73546 pEvol-pAzFRS.2.t1 pAzFRS.2.t1 E. coli p-azido-l-phenylalanine (pAzF) Bacterial..._AnapRS AnapRS E. coli 3-(6-acetylnaphthalen-2-ylamino)-2-amino-propanoic acid (Anap) Mammalian TAG Simon...Jesse Rinehart 71403 pCMV-DnpK PylRS M. barkeri N6‐(2‐(2,4‐dinitrophenyl)acetyl)lysine (DnpK) Bacterial,...pDule-IBBN (G2) IBBN (G2) synthetase M. jannaschii 4-(2′-bromoisobutyramido)-phenylalanine (IBBN) and structurally...pDule2-IBBN (G2) IBBN (G2) synthetase M. jannaschii 4-(2′-bromoisobutyramido)-phenylalanine (IBBN) and structurally...

  6. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...of basal H 2 O 2 levels with peroxiredoxin-based probes Real-time monitoring of basal H 2 O 2 levels with...Belousov Hydrogen Peroxide (H 2 O 2 ) Cytosolic or mitochondrial sensor for H 2 O 2 oxidation (roGFP2-Orp1) ...peroxiredoxin-2-based probe. Nat Commun. 2018 Aug 7;9(1):3145. Hadley Sikes Hydrogen Peroxide (H 2 O 2 ) Monitoring...encoded Ca(2+) indicator with enhanced two-photon absorption. Neurophotonics. 2024 Apr;11(2):024207. doi...Calcium Red fluorescent calcium sensors (fRCaMP1/2, GECO1/2) The kinetic mechanisms of fast-decay red-fluorescent...intracellular Zn(2+) homeostasis. Nat Methods. 2009 Oct;6(10):737-40. Maarten Merkx Zinc eZinCh-2 Zn2+ FRET ... eZinCh-2: A Versatile, Genetically Encoded FRET Sensor for Cytosolic and Intraorganelle Zn(2+) Imaging...

  7. Zhang Lab CRISPR Page

    Type
    Collection
    ...are also available. 2. SpCas9 (or SpCas9n, D10A nickase) + CRISPR RNA array + tracrRNA: This plasmid contains... system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9-Blast and lentiGuide-Puro...two MS2 RNA aptamers at the tetraloop and stemloop 2 The MS2-P65-HSF1 activation helper protein Full references...backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker. Contains BsmBI sites...inverted terminal repeats (ITR) from AAV serotype 2. SaCas9 only: This plasmid ( PX600, #61592 ) contains...Epub 2013 Aug 29. Erratum in: Cell. 2013 Oct 10;155(2):479-80. PubMed . Genome engineering using the CRISPR-Cas9... Feng G, Sharp PA, Zhang F. Cell . 2014 Oct 9;159(2):440-55. doi: 10.1016/j.cell.2014.09.014. Epub 2014...

  8. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...Dilute oligos 1:10 in ddH 2 O ( e.g., 1.0 μl annealed oligos + 9.0 μl ddH 2 O to yield a concentration...been included. Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation...), 50-100 ng gDNA, and H 2 O up to 20 μl. Use the primers designed in step 2 above. Conduct PCR for “non-deletion...have sufficiently proliferated once macroscopically visible with ~2 mm diameter. Primer Validation and ...frequency of indels formed on non-deleted alleles 2 , which are likely to cause frameshift mutations leading...of the Pim1 sgRNA from Table 1 are found in Table 2 . Obtain 24- or 25-mer oligos for each guide and its...sgRNA cleavage sites (“deletion band”; Figures 1 - 2 ). In the absence of deletion, the “deletion band”...

  9. Allen Institute for Brain Science AAV Enhancer Collection

    Type
    Collection
    ...Layer 2-3_IT Isocortex 230803 pAAV-AiE2638m-minBG-iCre(R297T)-BGHpA AiP20142 AiE2638m Cre Layer 2-3_IT ...Layer 2-3_IT Isocortex 220727 pAAV-AiE2543m-minBG-iCre(R297T)-BGHpA AiP2048 AiE2543m Cre Layer 2-3_IT ...Layer 2-3_IT Isocortex 230402 pAAV-AiE0680m-minBG-iCre(R297T)-BGHpA AiP15072 AiE0680m Cre Layer 2-3_IT ...plasmids for fluorescent labeling and recombination across brain regions and populations created by the Allen...specificity and brightness of fluorescent labeling across brain regions and populations using systemic AAV...based on their differential chromatin accessibility across different cell types and cloned into an AAV backbone...

  10. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...transcribed to make the pre-CRISPR RNA (pre-crRNA). (2) The pre-crRNA is processed into individual crRNAs...2015. Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain effector) Type I (Cas3) Type ... with unprecedented speed and specificity. Figure 2: An overview of CRISPR and NHEJ/HDR. The Cas9/gRNA...regulation of transcription in eukaryotes. Cell . 154(2):442-51. PMID: 23849981 Ishino Y, Shinagawa H, Makino...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell . 163(3):759-71. PMID: 26422227...trans-activating crRNA (tracrRNA) with homology to the short palindromic repeat. The tracrRNA helps recruit RNAse...together separate the individual crRNAs. (3) The tracrRNA and Cas9 nuclease form a complex with each individual...

  11. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ... achieved this by fusing the anti-CRISPR Acr protein, AcrIIA4, to a photo-sensitive LOV2 domain from A...the light-inducible heterodimerizing cryptochrome 2 (CRY2) and calcium and integrin-binding protein 1 ...inactive, dead Cas9 protein (dCas9) fused to CIB1; and 2) the activator - the CRY2 photolyase homology region... fused to both the N- and C-termini of dCas9; and 2) CRY2FL-VP64, a fusion of full-length CRY2 and the...researchers saw an 11-fold increase in mRNA levels after 2 hr and a 400-fold increase after 30 hr. The system...authors fragmented Cas9 into N-terminal (residues 2-713, N713) and C-terminal (residues 714-1368, C714...nMagC714 instead of nMagHigh1-C714, generating paCas9-2. This change did not significantly alter the system...

  12. Trimmer Lab NeuroMab Collection

    Type
    Collection
    ...FHF2 Human Mouse IgG2a 114527 Anti-JIP-2/IB-2 [N135/37.2R] JIP-2/IB-2 Mouse Mouse IgG2a 114529 Anti-Pan-SAPAP...Anti-CDY1/2/L1/L2 [N143/36R] CDY1/2/L1/L2 Human Mouse IgG2a 206535 Anti-CDY1/2 [N143/38R] CDY1/2 Human Mouse...206763 Neuroligin-2 scFv [L107/90] L107/90 scFv Neuroligin-2 Mouse Mouse 206764 Neuroligin-2 scFv [L107/95...receptor Rat Mouse IgG2a 114484 Anti-Stonin-2 [N346/9R] Stonin-2 Human Mouse IgG2a 114485 Anti-NSD3 [N348...Pan-FHF-A Human Mouse IgG2a 114510 Anti-REEP1/2 [N326D/29R] REEP1/2 Mouse Mouse IgG2a 114511 Anti-Kv3.1b K+ ...Mouse Mouse IgG2a 114547 Anti-Mitofusin-2 [N171/17.4R] Mitofusin-2 Mouse Mouse IgG2a 114548 Anti-Mortalin...Mouse IgG2a 140065 Anti-SCG10/Stathmin-2 [L5/1R] SCG10/Stathmin-2 Rat Mouse IgG2a 140066 Anti-Co-Rest/RCOR1...

  13. Optogenetics AAV Preps

    Type
    Collection
    ...Constitutive 1, 2, 5, 9 Deisseroth 26973 pAAV-hSyn-hChR2(H134R)-EYFP Syn ChR2/H134R EYFP Constitutive 1, 2, 5, 9....WPRE.SV40 CAG ChR2/H134R mCherry Constitutive 1, 2, 5, 9 Deisseroth 127090 pAAV-CAG-DIO-ChR2(H134R)-eYFP...dependent 1, 5, 8 Boyden 105669 pAAV-CKIIa-stGtACR2-FusionRed CaMKII GtACR2 (soma-targeted) FusionRed Constitutive...1, rg* Yizhar 105677 pAAV_hSyn1-SIO-stGtACR2-FusionRed Syn GtACR2 (soma-targeted) FusionRed Cre dependent...5, rg* Yizhar 105678 pAAV_hSyn1-SIO-stGtACR1-FusionRed Syn GtACR1 (soma-targeted) FusionRed Cre dependent...Yizhar 109048 pAAV-CAG-DIO-NLS-mRuby3-IRES-eGtACR1-ST CAG eGtACR1 (soma-targeted) mRuby3 Cre dependent 9 ...Inhibitory Arch and variants eNpHR and variants Jaws stGtACR iC++ OPN3 PdCO PPO Bidirectional BiPOLES Promoter...

  14. mTOR Pathway

    Type
    Collection
    ...known as RPS6KB1; Ribosomal protein S6 kinase B1 TSC1/2 TSC1 TSC2 Tuberous sclerosis VHL Von Hippel-Lindau...Rictor RPTOR independent companion of MTOR, complex 2 SGK Serum/glucocorticoid regulated kinase 1 Return...disease. Laplante M, Sabatini DM. Cell. 2012 Apr 13;149(2):274-93. doi: 10.1016/j.cell.2012.03.017. PubMed PMID...proliferation, including an abundant supply of macromolecules and energy, as well as the presence of both...

  15. Neurodegeneration Research Collection

    Type
    Collection
    ...for gene editing across divergent rodent species. Boender et al. Sci Adv. 2023 Jun 2. Use Cas9 in astrocytes...different inherited genes: Presenilin 1, Presenilin 2, and APP gene.The majority (>90%) of individuals develop...study aggregation. Saha et al. Nat Commun. 2023 Feb 2. Study the C terminal domain of TDP-43 to better understand...with the iPSC toolbox . Lam et al. bioRxiv. 2022 Dec 2. Use PiggyBac plasmids with tet-inducible expression...

  16. Viral Production

    Type
    Collection
    ...suggests that titers obtained using ddPCR are generally 2–3 fold higher than those achieved using the standard...catalog item (see maps section for images). Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre... a new vial of early passage cells. Approximately 2 weeks post-thaw, cell culture supernatant is tested...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...

  17. Cre-lox system

    Type
    Collection
    ...Cre-ERT2 CAG Mammalian Heller 112622 pVHC_PGKneoLox2DTA.2 Venus, Cre-ERT2, targeting vector with MCS for homology...homology arms Mammalian Heller 112623 pEHC_PGKneoLox2DTA.2 Emerald, Cre-ERT2, targeting vector with MCS for homology... arms Mammalian Heller 112624 pmTHC_PGKneoLox2DTA.2 TFP, Cre-ERT2, targeting vector with MCS for homology...arms Mammalian Heller 112625 ptdTHC_PGKneoLox2DTA.2 tdTomato, Cre-ERT2, targeting vector with MCS for ...25;1504(4):467-86. doi:10.1016/0022-2836(81)90375-2. PubMed . Ventura A, Meissner A, Jaenisch R, Jacks...photo-activatable cre recombinase CAG AAV Bruchas 78398 pNXRVa-HACre Cre CMV Yeast Tomita 82587 pME-ERT2-Cre-ERT2 Tamoxifen-inducible...Cre-PGK-Puro Cre EF-1 alpha Lentiviral Alcudia 108639 fli1aCreERT2 Cre-ERT2; endothelial reporter fli1a Zebrafish...

  18. CRISPR References and Information

    Type
    Collection
    ...Link opens in a new window) An encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro...and BsmBI site for cloning in gRNA: pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) PDF, 49 KB Vosshall...RNA array: Cas9 (pX260) or Cas9 D10A (pX334) ; tracrRNA: Cas9 (pX330) or Cas9 D10A (pX335) PDF, 255 KB...

  19. Bikard Lab - CRISPR Repression Collection

    Type
    Collection
    ...chromosomal integration of DNA. ACS Synthetic Biology, 2(9), 537–541. DOI: 10.1021/sb400021j Figure 1: Multiplexing...repeat motifs (squares), and also carrying the tracrRNA sequence (not shown). (B) Relative GFP and RFP...

  20. Brain Initiative Collection

    Type
    Collection
    ...Gordon Fishell 83899-AAV1 pAAV-mDlx-GCaMP6f-Fishell-2 GCaMP6f expression in forebrain GABA-ergic interneurons...Gordon Fishell 83899-AAV9 pAAV-mDlx-GCaMP6f-Fishell-2 GCaMP6f expression in forebrain GABA-ergic interneurons...Gordon Fishell 83899-AAVrg pAAV-mDlx-GCaMP6f-Fishell-2 GCaMP6f expression in forebrain GABA-ergic interneurons...AAV9 pAAV-CAG-DIO-NLS-mRuby3-IRES-eGtACR1-ST Anion channelrhodopsin GtACR1 fused to ER export signal and ...
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