...pcDNA3-EGFP - Mammalian Expression pCAG-GFP - Mammalian Expression pLV-eGFP - Mammalian Lentiviral Expression...A206K) pmEGFP-1 - Mammalian Expression mEGFP-N1 - Mammalian Expression mEGFP-C1 - Mammalian Expression mEGFP-pBAD...pCAG-DsRed2 - Mammalian Expression DsRed2-N1 - Mammalian Expression DsRed2-C1 - Mammalian Expression DsRed2...pCAG-HcRed - Mammalian Expression HcRed1-N1 - Mammalian Expression HcRed1-C1 - Mammalian Expression dKatushka...pQC mPlum IX - Mammalian Expression mPlum-N1 - Mammalian Expression mPlum-C1 - Mammalian Expression mPlum-pBAD... Sirius-N1 - Mammalian Expression SBFP2 380 446 13 Monomer (A206K) pSBFP2-C1 - Mammalian Expression Azurite...Monomer (A206K) mAzurite-N1 - Mammalian Expression mAzurite-C1 - Mammalian Expression EBFP2 383 448 17 ...

...gRNA_Cloning Vector 41824 Mammalian AfIII none S. pyogenes Bleocin Church MLM3636 43860 Mammalian BsmBI none S. pyogenes...pSPgRNA 47108 Mammalian BbsI none S. pyogenes Gersbach pUC57-sgRNA expression vector 51132 Mammalian BsaI none...sgExpression 48240 Mammalian BbsI yes, activate S. pyogenes Jaenisch pSQT1313 53370 Mammalian BsmBI none S....Gen) 48140 Mammalian BbsI yes, nick S. pyogenes EGFP Zhang PX460 (3rd Gen) 48873 Mammalian BbsI yes, nick... Gen) 48138 Mammalian BbsI yes, cut S. pyogenes EGFP Zhang PX335 (2nd Gen) 42335 Mammalian BbsI yes, nick...Gen) 42333 Mammalian BbsI yes, nick S. pyogenes Puro Zhang PX330 (2nd Gen) 42230 Mammalian BbsI yes, cut...42229 Mammalian BbsI yes, cut S. pyogenes Puro Zhang sgRNA(MS2) cloning backbone 61424 Mammalian BbsI ...

...cTnT Mammalian Campbell 89573 pCAG-iCre iCre CAG Mammalian Wong 89575 pCAG-VCre VCre CAG Mammalian Wong... CAG Mammalian Scholer 125574 pCAG-Cre Cre CAG Mammalian Imai 125577 pCAG-iCre iCre CAG Mammalian Imai...p209 pCMV-cre-K Cre-K CMV Mammalian Green 8395 p210 pCMV-CREM CREM CMV Mammalian Green 8401 p224 pCMV-RFP...floxed RFP within CREM CMV Mammalian Green 8403 p153 pPr-cre Cre probasin Mammalian Green 8404 p236 pPr-CREM...CREM probasin Mammalian Green 8405 p240 pPr-CREM/Ins CREM with insulators probasin Mammalian Green 8406 ...separated by insulator probasin Mammalian Green 11543 pPGK-Cre-bpA Cre PGK Mammalian Rajewsky 11916 pBS185 CMV-Cre...CMV-Cre Cre hCMV Mammalian Sauer 11918 pBS513 EF1alpha-cre Cre EF-1 alpha Mammalian Sauer 11919 pBS448...

...(Anl) Mammalian ATG David Tirrell 64915 pMAH-POLY tyrosyl-tRNA synthetase E. coli pBof Mammalian TAG Huiwang...AbK Mammalian TAG Simon Elsaesser 140011 pAS_4xMx PylT_FLAG-Mx PylRS PylRS M. alvus CpK Mammalian TAG ...-aminotyrosine Mammalian Huiwang Ai 154762 pAS_FLAG-Mma PylRS PylRS M. mazei Mammalian TAG Simon Elsaesser...archaeon Mammalian TAG Simon Elsaesser 154764 pAS_FLAG-Mx1201 PylRS PylRS M. alvus Mammalian TAG Simon...acetyl-lysine Mammalian Tao Liu 164081 pCMV-MfPylRS (human-opti) PylRS M. flavescens acetyl-lysine Mammalian Tao...mazei Mammalian TAG Simon Elsaesser 174896 pAS_4xMmaPylT(UUA)_EF1_MmaPylRS PylRS M. mazei Mammalian TAA ...mazei Mammalian TGA Simon Elsaesser 174901 pAS_MmaPylT_EF1_NES-MmaPylRS WT PylRS M. mazei Mammalian TAG ...

...3xHA tag for mammalian expression pHAHA - N-terminal double HA tag for mammalian expression...pMKO.1 puro - Mammalian retroviral shRNA expression MSCV-IRES-GFP - Mammalian ...plasmid for mammalian gene silencing pLVTHM - lentiviral plasmid for mammalian gene silencing...Relevant Promoters Representative Empty Backbones Mammalian CMV, SV40, EF1a, CAG pSG5L Flag HA - Transient... expression - Gradia Lab plasmids for mammalian expression, Ligation Independent...Flag HA - N-terminal Flag-HA for mammalian expression pNIC-CTHF - C-terminal ...pKMyc - N-terminal Myc tag for mammalian expression 6xHis Epitope Tag pEZYmyc-His - C-terminal...

...pAquaN1 Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express a gene...subcellular compartments pCEP4CyPet-MAMM Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial...Yellow Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-terminal...Yellow Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover Green Mammalian Expresses...Pre-constructed fluorescent biosensors targeting small molecules or specific genes are also available. ...N-terminus of monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus...C-terminus of monomeric Cerulean mTurquoise2 Cyan Mammalian Constructs to target mTurquoise2 to various subcellular...

...promoter for normalization. FMDV StopGo sequences flank the MCS for improved normalization. John Atkins... CMV Mammalian expression of Nano-lantern Takeharu Nagai 87121 pcDNA-RLuc8 Renilla CMV Mammalian expression...of organisms, ranging from bacteria to aquatic animals to insects. Luciferase catalyzes the oxidation ...that has been codon optimized for expression in mammalian systems. Other luciferases in our collection include...has also been codon optimized ( CBR2opt ) for mammalian expression systems. Each type of luciferase has...due to their large size. NanoLuc® Luciferase is small, bright, and a secreted version of the protein is...protein interactions , protein dimerization , endosomal escape of cell-penetrating peptides , and more...

... 293 Mammalian (pcDNA3) GAP ELMOD3 84173 381 Mammalian (pcDNA3) GEF Arfgef1 10565 1849 Mammalian (pcDNA4c...Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase Arf3 377 181 Bacterial (pET), Mammalian (pcDNA3), Gateway...Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase Arf5 381 180 Bacterial (pET), Mammalian (pcDNA3), Gateway...Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase Arl3 403 182 Bacterial (pET), Mammalian (pcDNA3), Gateway...1704 GAP Arap3 64411 1544 Mammalian (pEGFP-C2) GAP Acap1 9744 740 Mammalian (pFLAG-CMV2) GAP Acap2 23527...compatible GEF, GTPase, and GAP set is referred to as a minimal triad, with each component involved in GTPase signaling...Gateway GTPase Arf6 382 175 Bacterial (pET), Mammalian (pcDNA3), Gateway GTPase Arl1 400 181 Bacterial ...

...Browse Plasmids: Anti-CRISPR Small Precision Edits CRISPR can be used to make small, very precise edits of ...double-stranded DNA plasmid. Small ssDNA oligos often work well for small edits like a single-point mutation...Guide Jump To: CRISPR Guide Overview CRISPR Basics Small Edits Large Edits Genome-Wide Screening Cas9 Fusion...missing sequence, and is more commonly used for small precision edits. NHEJ directly ligates the break...efficient in repairing DSB, it frequently causes small nucleotide insertions or deletions (indels) at the...modification ). In most cases, NHEJ gives rise to small indels in the target DNA that result in amino acid... handful of bases, or even a single base. These small precision edits can be done via HDR, base editing...

...methods: in animals, in cultured immune cells, or through the use of recombinant plasmids. Animals - Antibodies...Antibodies produced in animals make use of the animal’s immune system. The antigen of interest is introduced...introduced to the animal, which then mounts an immune response to it. Once the immune response has developed...antibody. Monocultures of B cells isolated from animals immunized as described above are used to create... protein chains: two large heavy chains and two small light chains. These four domains come together to...only produce one variable region. Isotypes The mammalian immune system produces five classes of antibodies...antibodies are typically collected directly from animals and contain antibodies targeting various epitopes...

...29;7:12266. David Savage Maltose Monitor maltose levels and metabolism (FLIPmal) Fluorescence Resonance...:19562. Takashi Tsuboi Maltose Single fluorescent protein biosensor for maltose Rapid construction of ...Jun 3. 1(1):11. Wolf Frommer Maltose Green fluorescent MBP-based maltose biosensor A genetically encoded...Biosensors Fluorescent Proteins: Biosensors Metal Ions Small Molecules Neurotransmitters Cell Physiology Browse...Diverse Functional Payloads to Cells of Behaving Mammals. Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-...tethering by PDZD8 regulates Ca(2+) dynamics in mammalian neurons. Science. 2017 Nov 3;358(6363):623-630...

...available for expression in mammalian systems, bacteria, plants, and Drosophila. Mammalian ID Plasmid Gene/Insert...GG-acceptor Mammalian hU6 epegRNA BsaI No mRFP1 David Liu 174039 pU6-tmpknot-GG-acceptor Mammalian hU6 epegRNA...Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish... activity; can be used in PE2–PE5 systems PE6 - small prime editors with optimized RT and/or Cas9 domains...Cas9 Selection PI 132777 pU6-pegRNA-GG-acceptor Mammalian hU6 pegRNA BsaI No mRFP1 David Liu 140448 QPM-... No Vermilion Norbert Perrimon 164423 pHSG1C3 Mammalian U6 pegRNA + nicking sgRNA BbsI for sgRNAs; BbsI...sgRNA No Tilmann Weber 173220 pPBT-peRNA_GG-Puro Mammalian, piggyBac hU6 pegRNA BsmBI No Puromycin Jacob ...

...3xFNLDD in mammalian cells 48880 3xFNLDD/pMXs-I2 iChIP plasmid, expresses 3xFNLDD in mammalian cells 48972...3xFNLDD in mammalian cells 49536 3xFNLDD/pMXs-puro iChIP plasmid, expresses 3xFNLDD in mammalian cells 51240...7.1 enChIP plasmid, expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic...3xFLAG-dCas9/pMXs-puro Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic...3xFLAG-dCas9/pMXs-IG Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic...3xFLAG-dCas9/pMXs-I2 Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic...3xFLAG-dCas9/pMXs-neo Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic...

...Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or gRNA in a mammalian cell line...Although we will use the example of CRISPR/Cas9 in mammalian cells, many of these principles apply to using...dCas9-KRAB is more effective than dCas9 alone for mammalian cell lines. Activate (CRISPRa) Increase expression...expression systems and variables for using CRISPR in mammalian cells. Some of the variables include: Species ...Expression System Components of System Application Mammalian expression vector Cas enzyme promoter can be constitutive...expression of Cas9 and/or gRNA in a wide variety of mammalian cell lines Useful for difficult to transfect cell... ∼4.5 kb packaging limit (only compatible with smaller Cas enzymes) Transient or stable expression of ...

...-Destination constructs were designed for optimal mammalian expression of Voytas Golden Gate-assembled...destination vectors and a reporter vector for mammalian cell-based validation assay that are designed ...vectors, pcDNA-TAL-NC2 and pCAGGS-TAL-NC2, are mammalian expression/mRNA synthesis vectors containing unique... contains the VP64 domain (four repeats of the minimal activation domain of VP16) at the C terminus. 47389...Torres-Padilla These plasmids were designed for mammalian expression of fluorescent TAL effectors to visualize...plasmids were optimized for TALEN expression in mammalian systems including hard-to-transfect cell lines...sequence to drive efficient TALEN expression in mammalian cells, (iv) an improved, truncated TALE backbone...

...Bacterial systems were tested for functionality in mammalian cells. The lac system with the inducer isopropylthio-β-galactoside... Gossen and Bujard tested the tet system in a mammalian cell system (HeLa) and found that the tet system...promoters are developed by placing a TRE upstream of a minimal promoter. Tetracycline Off System The initial system...promoter (Pbi); Pbi contains a TRE between two minimal CMV and is silent in absence of binding of tTA ...knockdown of gene expression at a single copy in mammalian cells; Expresses firefly luciferase hairpin and...References Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Gossen...

...affecting females at an incidence of approximately 1:10,000. It is characterized by apparently normal development...Syndrome Plasmid Collection and Resource Center MECP2 Animal Models Human Cell Lines CRISPR Tools Plasmids Resources...criteria, defined as the presence of relatively normal early development, regression with a loss of spoken... rare cases Rett syndrome can be inherited. In females, random X inactivation results in approximately...PMID: 21074045 2017 - the Bird lab determines a minimally functional MECP2 that rescues neurological defects...opens in a new window) PMID: 29019980 Return to top Animal Models Studies of the MECP2 protein in mouse and...general introduction to CRISPR technology or the mammalian CRISPR resources for a full selection of plasmids...

... nuclease domains: pST1374 (for expression in mammalian cells or zebrafish) pDW1775 (for expression in... domain. Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute...domain ( Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish). pMLM802 (for expression of...domain ( Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish). pMG10 (a '“2-in-1” ZFN cassette...domain ( Sollu et al., Nucleic Acids Res ) in mammalian cells or zebrafish). pCLR2068 (for expression ...domain. ( Doyon et al., Nat Methods 2011 ) in mammalian cells or zebrafish as described by Moore et al...

...2013, Cong et al. and Mali et al. described CRISPR-based genome editing in mammalian cell culture. Five ... specialized Cas proteins snip foreign DNA into small fragments approximately 20 bp in length and paste...characterized. Staphylococcus aureus (SaCas9) is smaller than SpCas9 and more easily packaged in AAV. Cas9... improving targeting in AT-rich genomes. Cpf1's small size also makes it suitable for multiplexing and...permanent genome edits or double-strand breaks. In mammalian cells, Cas13-ADAR2 does not require a specific... this non-specific cleavage does not occur in mammalian cells. Purify : Epitope-tagged dCas9 can also ...has CRISPR plasmids for the following models: Mammalian systems (Human, mouse, and rat) Bacteria ( E. ...

...are sporadic, having no prior family history. A small percentage (5-10%) are familial ALS cases having...These familial cases, usually inherited in an autosomal dominant manner, are associated with known genetic... Disease Huntington’s disease (HD) is a fatal autosomal dominant genetic disorder that causes the progressive...difficulty speaking and swallowing. HD is caused by abnormal expansion of a repeating CAG triplet series within...mutated HTT gene will have huntingtin proteins with abnormally long polyglutamine sequences. These glutamine-rich...medication and surgery. PD primarily involves the malfunction and death of dopamine-producing neurons in the... specific known cause) nature of the disease, a small portion of cases can be attributed to known genetic...