...immunoglobulin heavy diversity 3-22 IGHD322 IGHD3-3 immunoglobulin heavy diversity 3-3 DXP4, IGHD33 IGHD3-9 immunoglobulin...VH IGHV3-30-3 immunoglobulin heavy variable 3-30-3 IGHV3-3, IGHV3303 IGHV3-30-5 immunoglobulin heavy variable...motif) ligand 3-like 2 G0S19-3, LD78gamma, SCYA3L2 CCL3L3 chemokine (C-C motif) ligand 3-like 3 464.2, D17S1718... variable 3-30-5 IGHV3-3, IGHV3305 IGHV3-33 immunoglobulin heavy variable 3-33 IGHV333, VH IGHV3-35 immunoglobulin...lambda variable 3-9 (gene/pseudogene) IGLV39, V2-6 IGLV4-3 immunoglobulin lambda variable 4-3 IGLV43, V5-1..., MRS DEFA3 defensin, alpha 3, neutrophil-specific DEF3, HNP-3, HNP3, HP-3 DEFA5 defensin, alpha 5, Paneth...

...Human Parts 3rd 3 59,576 Human Whole Genome sgRNA iBAR Library 140633 Knockout Human Wei 3rd 3 58,630 Human...SAM v1 - 3 plasmid system 1000000057 (Zeocin) 1000000074 (Puromycin) Activation Human Zhang 3rd 3 70,290...70,290 SAM v1 - 3 plasmid system 1000000075 (Puromycin) Activation Mouse Zhang 3rd 3 69,716 SAM v2 - 2 plasmid...177663, 177664, 177670 Inhibition Human Adamson 3rd ∼3 366 or 1,573 Activity-optimized genome-wide library...92380 (Set B) Activation Human Doench and Root 3rd 3–6 56,762 (Set A) 56,476 (Set B) Broad GPP activation...92384 (Set B) Activation Mouse Doench and Root 3rd 3–6 67,187 (Set A) 66,889 (Set B) Broad GPP inhibition...92386 (Set B) Inhibition Human Doench and Root 3rd 3–6 57,050 (Set A) 57,011 (Set B) Broad GPP inhibition...

...RLD-1 nuclear receptor subfamily 1, group H, member 3 NR2E1 Plasmids NR2E1, RP3-429G5.1, TLL, TLX, XTLL ...group E, member 1 NR2F1 plasmids NR2F1, COUP-TFI, EAR-3, EAR3, ERBAL3, NR2F2, SVP44, TCFCOUP1, TFCOUP1 nuclear...NR3C1, GCCR, GCR, GR, GRL nuclear receptor subfamily 3, group C, member 1 (glucocorticoid receptor) NR3C2...MCR, MLR, MR, NR3C2VIT nuclear receptor subfamily 3, group C, member 2 NR4A1 Plasmids NR4A1, GFRP1, HMR...subfamily 4, group A, member 1 NR4A2 Plasmids NR4A2, HZF-3, NOT, NURR1, RNR1, TINUR nuclear receptor subfamily...nuclear receptor plasmid collection assembled in collaboration with the Nuclear Receptor Signaling Atlas (NURSA...associated with numerous disease pathways. In collaboration with research labs from around the globe, Addgene...

...EQR variant 3' NGAG SpCas9 VQR variant 3' NGAN or NGNG xCas9 3' NG, GAA, or GAT SpCas9-NG 3' NG Staphylococcus...SP); SpCas9 3' NGG SpCas9 D1135E variant 3' NGG (reduced NAG binding) SpCas9 VRER variant 3' NGCG SpCas9...jejuni (CJ) 3' NNNNRYAC Neisseria meningitidis (NM) 3' NNNNGATT Streptococcus thermophilus (ST) 3' NNAGAAW...characterized *In the table above, 3' and 5' indicate on which end of targeted sequence the PAM is located... DNA target, the seed sequence (8–10 bases at the 3′ end of the gRNA targeting sequence) will begin to...gRNA will continue to anneal to the target DNA in a 3′ to 5′ direction. The location of any potential mismatches...matters. Mismatches between the target sequence in the 3′ seed sequence inhibit target cleavage, while mismatches...

...with four different phosphobinding domains (14-3-3β, 14-3-3σ, the WW2 domain of NEDD4, and the WW2 domain...111881 pNAS1b split mCherry 14-3-3σ Plasmid 111880 pNAS1b split mCherry 14-3-3β Pooled Library 111704 Mode...111705 Mode #2 Library (14-3-3β) Pooled Library 111706 Mode #2 Library (14-3-3σ) Pooled Library 111707 ...iSPI_pSer_Subpool#2 Pooled Library 188528 iSPI_pSer_Subpool#3 Pooled Library 188529 iSPI_pSer_Subpool#4 Pooled Library...52055 EcAR7 Plasmid 112031 pNAS1b split mCherry 14-3-3β ctrl Plasmid 112030 pNAS1b split mCherry NEDD4 ...This library is anticipated to be useful for laboratories performing phosphoproteomic experiments in need...

... AKT3 v-akt murine thymoma viral oncogene homolog 3 ALK Anaplastic lymphoma receptor tyrosine kinase ARHGAP35... factor receptor FLT3 Fms related tyrosine kinase 3 FNT FNTA FNTB Farnesyltransferase, CAAX box FOS FBJ...PDGFRB Platelet derived growth factor receptor PDPK1 3-phosphoinositide dependent protein kinase 1 PEBP1 ...known as PI3K; Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic and regulatory subunits - Class I...cancer therapy. Downward J. Nat Rev Cancer. 2003 Jan;3(1):11-22. PubMed PMID: 12509763 . Do you have suggestions... Frederick National Laboratory for Cancer Research . Frederick National Laboratory for Cancer Research...

...sequence in the 3' UTR David Bartel 12179 pIS1 Renilla Insertion of target sequence in the 3' UTR David Bartel...the middle of the Firefly luciferase gene.. TREAT (3( T hree)′- R NA E nd A ccumulation during T urnover... pIS2 Renilla Insertion of target sequence in the 3' UTR David Bartel 101139 pCMV-IRES-Renilla Luciferase-IRES-Gateway-Firefly...Firefly Insertion of imperfect miRNA sequence in the 3' UTR Carl Novina 33154 pAC-Luc Firefly Creation of...Atkins 167576 mpCHECK2 Firefly/Renilla Insertion of 3' UTR sequences downstream of Renilla luciferase. Firefly...Macho 136010 psi-CHECK3 Firefly/Renilla Insertion of 3' UTR sequences downstream of Renilla luciferase. Firefly...Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016 blog post...

...homolog GSK3 GSK3A GSK3B Glycogen synthase kinase 3 MDM2 MDM2 proto-oncogene mSin1 Also known as MAPKAP1...PIK3R5 PIK3R6 Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic and regulatory subunits - Class I...Li J, Kim SG, Blenis J. Cell Metab. 2014 Mar 4;19(3):373-9. doi: 10.1016/j.cmet.2014.01.001. PubMed PMID... the cell and regulates the switch from anabolic to catabolic processes. Active mTORC1 promotes protein...mTOR pathway. enhanced signaling of mTOR, a key metabolic regulator, correlates with poor prognosis in many...ammalian t arget o f r apamycin (mTOR) is a key metabolic regulator controlling cell growth and proliferation...energy-intensive processes. mTORC1 also blocks catabolic processes including autophagy and lysosome biogenesis...

...Lentiviral reverse tetracycline-controlled transactivator 3 (rtTA3) expression vector, CMV promoter, and Blasticidin...tTA at its 5' end and an inducible promoter at its 3' end, which controls GFP expression tTA Off Verma ...contains insulator sequence; plasmid pEnt L1L3 rtTA-3 is a rtTA version tTA Off Hsiao 34856 fos-tTA Mouse...plus part of the first intron (-764/+918) and fos 3'UTR tTA Off Mayford 38056 pXCX.CMV.tTA Adenoviral;...Oesch F, Zabel B. Physiol Genomics . 2002 Dec 3;11(3):115-32. PubMed . Tetracycline derivatives: alternative... or shRNA; see article for detailed information about cloning and additional plasmids tetR-KRAB On Aebischer...

...Anti-Synaptotagmin-3 [N278/19R] Synaptotagmin-3 Mouse Mouse IgG2a 114477 Anti-TRPML3/Mucolipin-3 [N268/19R] TRPML3... victoria Mouse IgG2a 114493 Anti-NGL-3/LRRC4B [N51/6.2R] NGL-3/LRRC4B Rat Mouse IgG2a 114494 Anti-VGlut3...Rat Mouse IgG2a 114518 Anti-Neuroligin-3 [N110/29.4R] Neuroligin-3 Rat Mouse IgG2a 114519 Anti-RGS14 [N133...149452 Anti-Kirrel3, short isoform 3 [N320/48R] Kirrel3, short isoform 3 Mouse Mouse IgG2a 149453 Anti-Kv4.2...Bassoon Rat Mouse IgG2a 177471 Anti-Synapsin-3 [L125/121R] Synapsin-3 Human Mouse IgG2a 177472 Anti-Pan-Synapsin...Mouse Mouse IgG2a 206608 Anti-Synapsin-3 [L125/76R] Synapsin-3 Human Mouse IgG2a 206610 Anti-Cav1.3 Ca2...Mouse Mouse IgG2b 206652 Anti-Neuroligin-3 [N110/29R-2b] Neuroligin-3 Rat Mouse IgG2b 206653 Anti-HCN4 [N114...

...pX330 vector using BbsI restriction enzyme ( Table 3 ). Add a G nucleotide after the CACC sequence and ...nucleotide after the CACC sequence. Add a C at the 3’ end of the reverse complement oligo ( e.g., sgRNA-A...pX330 using a Golden Gate assembly cloning strategy 3 . Prepare a 50 μl reaction mix: 100 ng circular pX330...ml ampicillin and incubate O/N at 37 °C. Pick 2 - 3 colonies and inoculate into a mini-prep culture. Perform...50 μm filter into a FACS tube. FACS sort the top ~3% of GFP positive cells in order to enrich for cells...plate. Allow the bulk cells to incubate at 37 °C for 3 - 7 days and allow the clones to incubate at 37 °C...article from Journal of Visualized Experiments (JoVE) about genomic deletions in mammalian cell lines using ...

...Barcode Library Version 3 Ready-to-use lentiviral particles carrying version 3 of the CellTag barcoding...barcoding library. Version 3 of the Celltag library contains 5737 barcodes to combinatorially index cells for...are reported with each shipment. For more details about our viral vector packaging service, visit our Viral...

...Mammals. Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. Karl Deisseroth , INTRSECT 2.0 Project Calcium Expression...Mammals. Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. Karl Deisseroth , INTRSECT 2.0 Project Calcium Mitochondrial...Chromosome Condensation. Curr Biol. 2018 Feb 5;28(3):444-451.e6. Takeharu Nagai Magnesium MagFRET-1 Mg2...During Nutrient Transitions. ACS Sens. 2020 Mar 27;5(3):814-822. Bas Teusink ATP (extracellular) ecAT3.10...accumulation in bacteria. Biotechnol Biofuels. 2008 Jun 3. 1(1):11. Wolf Frommer Maltose Green fluorescent MBP-based...Learning. Cell. 2020 Dec 12. pii: S0092-8674(20)31612-3. Lin Tian Serotonin Serotonin (5-HT) sensor psychLight...biosensor. Cell. 2021 Apr 8. pii: S0092-8674(21)00374-3. Lin Tian Serotonin Cilia-targeted serotonin sensor...

...) and need to be followed on the 3' end by a 3bp NGG PAM sequence. 3. SpCas9 alone: This plasmid contains...site sequence (20bp) and need to be followed on the 3' end by a 3bp NGG PAM sequence. We have found that...Targeted gene activation using SAM There are two sets of 3 vectors each available for mammalian endogenous gene...sequence (21-22bp) and need to be followed on the 3' end by NGRRT or NGRRN PAM sequences. We have found...Jiang W, Marraffini LA, Zhang F. Science . 2013 Jan 3. DOI: 10.1126/science.1231143. PubMed . DNA targeting...Addgene Zhang Lab CRISPR Resources Tips for and FAQs about the Zhang Lab CRISPRs Browse All Zhang Lab Plasmids...

...tyrosine-mutant AAV serotype vectors. Mol Ther . 2009 Mar;17(3):463-71. PMID: 19066593 Other citations include: Bogner...cell entry characteristics. Gene Ther . 2020 Apr;27(3-4):127-142. PMID: 31611639 AAV2(trpYF) When using ...Expressing Retinoschisin. Hum Gene Ther Clin Dev . Sep;26(3):165-76. PMID: 26390090 Other citations include: Scalabrino...Information The plasmid below is available in each of the above serotypes and buffers. Please click on the plasmid...

... actual oligo you should order is: 5'-CACCGN...N-3' 3'-CN...NCAAA-5' You need to add the CACC and AAAC...region, will they also be targeted? The Cas9 cuts 3-4bp upstream of the PAM sequence. There can be some...)01015-5 As for target selection, we usually pick 3-6 guides around the region to find the most efficient... Feng Zhang's Lab, has assembled a list of FAQs about using the lab's CRISPR technology. Genome..., February 2013 ), has assembled a list of FAQs about using the lab's CRISPR technology. Visit the Zhang...

...inducible (Tet) expression miRNA and 3' UTR mRNA regulation pIS0 - 3'UTR segments of target genes can be... for gene expression Thy1.1 Mammalian pLKO.3 Thy1.1 - Lentiviral shRNA expression Gentamicin Varies...see more Gateway vectors) pCAP03-acc(3)IV - Facilitate the capture and expression of bacterial... your gene and of the fusion protein. Read more about epitope tags and protein tags . Tag or Fusion ...functions such as protein purification mentioned above, empty backbones can also be used for for viral ...our dedicated Cre-lox Plasmids page pBF3060 - Metabolic engineering in S. cerevisiae TALENs Gene targeting...

...development (embryogenesis is complete in ~3 days), short generation time (~3 months), ease of care and breeding...spectral barcodes for cell tracing - Sean Megason Lab Zebrabow - Albert Pan and Alex Schier Labs. Multicolor ...zebrafish strains and materials and information about zebrafish research. China Zebrafish Resource Center...America. The Zebrafish Book - A guide for the laboratory use of zebrafish. CRISPRz - A database of CRISPR...gene expression database that provides information about when genes are active in developing embryos. zfRegeneration...

... that titers obtained using ddPCR are generally 2–3 fold higher than those achieved using the standard... appear darker than full vector particles. Figure 3: Electron micrograph of AAV vector preparation shows... before being distributed to customers. Details about our production protocols, titering methods, and ...either in-house by Addgene scientists or through collaboration with viral vector manufacturing facilities, ...

...(exo-) DNA Polymerase I, Large (Klenow) Fragment (3'-5' exo-) 165512 pOpen-THEAQpolA Taq DNA Polymerase...MMLV) Reverse Transcriptase (RNAse H deactivated by 3 mutations) 165556 pOpen-MMLV_RT (lack H) Moloney Murine...engineering and directed evolution. More information about the collection can be obtained at the Open Bioeconomy...

...p38Beta MAPK12 ERK3, ERK6, P38GAMMA, PRKM12, SAPK-3, SAPK3 MAPK13 RP1-179N16.4, MAPK 13, ... point for multiple biochemical signals. In collaboration with numerous research labs from around the ...

...of bacterial genes. Molecular Systems Biology, 14(3):e7899. doi: 10.15252/msb.20177899. PMID: 29519933...attachment site of the phage 186. Detailed information about the repression system can be found in this article...29519933 . bioRxiv preprint . Detailed information about chromosomal integration of pLC97 can be found in this...

...). DeChenne-Peters et al. PLoS One. 2023 Mar 9;18(3):e0282170. PMID: 36893201 (Link opens in a new window...integrating MDH-related research into their teaching laboratories. They encourage instructors teaching different...research community and facilitates scientific collaborations between institutions. With these plasmids, ...regulation and post-translational modifications, and metabolic pathways and protein interactions. Plasmids A ...projects, find teaching resources and protocols, learn about regional hubs, access a teaching mentor, and more....2021.43 (Link opens in a new window) External Collaboration Results in Student Learning Gains and Positive...

...Transfer plasmids with sequence from the 5′ to the 3′ ITR exceeding 4.7 kb Transfer plasmids with a cargo... by email will supersede the general guarantees above. We will have our expert viral vector production...supersede the general estimated turnaround time above. After starting viral production, if we determine...

... consists of 5 individually packaged plasmids and 3 individually packaged strains, which are shipped as...Dove et al., Nature 1997 ) and later modified by Pabo and colleagues ( Joung et al., PNAS 2000 ). The ...

... work in your mouse line, see Supplementary Table 3 (Link opens in a new window) . Browse Available PHP.eB...Inhibitory - AAV-PHP.eB with GRE or transgenic animal L2/3 & L4 - AAV.CAP-B10 with GRE or transgenic animal Excitatory...systemic serotypes from the Viviana Gradinaru laboratory and the Caltech CLOVER Center (Link opens in ...AAV Decision Tree The AAV decision tree graphic above helps you choose the best capsid for your cell type...

...Combinatorial Strategies Connie Cepko 2:45 PM Break 3:00 PM (Group Discussion) Technologies and Resources...Learn about Addgene's 2018 AAV Education and Development (AAVED) workshop series. The... Institute Katherine Matho Cold Spring Harbor Laboratory Gowan Tervo Janelia Research Campus Sarada Viswanathan...Institute Katherine Matho, Cold Spring Harbor Laboratory Gowan Tervo and Sarada Viswanathan, Janelia Research...

... rpoZ- on plates lacking histidine and containing 3-AT Scot Wolfe Bacterial one-hybrid system for testing...promotes growth on plates lacking His and containing 3-AT. FLIParaF.Ec-250n 18855 N/A FRET between CFP and...in vitro ? These, in addition to many questions about bacteria themselves, can be answered using model...overexpressing or knocking down a gene can tell you much about its function and/or alter cellular activity in a...basic biology of a bacterial gene or redirecting metabolic pathways to produce a therapeutic compound, there...once you get it right, you can learn many things about your protein’s function, structure, and interactions...molecules, light, temperature, or even intracellular metabolite levels. Different plasmids and, often times, ...

...genetic model organism due to their rapid life cycle (3 days from egg to adult) and easy lab maintenance. ... approach has also been developed by the Fire laboratory and described in Efficient Marker-Free Recovery...

...Cre lac Bacterial Ekker 47551 pDD104 (Peft-3::Cre) Cre eft-3 C. elegans Goldstein 48201 CAG-GFP-IRES-CRE...deltaG)-Cre Cre CMV RABV Jessell 73717 pMLS328 Cre eft-3 C. elegans Jorgensen 75267 pAAV-CIBN-CreC photo-activatable...Vivo Electroporation. Proc Natl Acad Sci. Jan 16;104(3):1027-32. doi: 10.1073/pnas.0610155104. Epub 2007 ...Learn about the Cre-Lox system and how it can be used for genetic rearrangements and recombination. Use...

...Fishell 83898-AAV1 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...Fishell 83898-AAV9 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...Fishell 83898-AAVrg pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...particles that are suitable for in vivo use. Learn more about our viral service ID Viral Prep Description PI 65417...well as by our trusted partner labs. Learn more about our antibodies ID Recombinant Antibody Reactivity...

...TALEs for the masses. Rusk N. Nat Methods 2011 Mar;8(3):197. Pubmed PMID: 21473016 . Move over ZFNs. Nat ...researchers who are learning to wield them. Learn more about Addgene’s TAL effector and TALEN kits here . References...

...standard consisting of ECFP and EYFP connected by 3 GGSGGS repeats pET28CLY4 Peptide linker standard consisting...constructed using the cpFRET kit from the Pertz laboratory. Empty Vectors Encoding Fluorescent Proteins ...

...shRNA cassettes into the unique SnaBI site in the 3´-LTR). See here for other versions of pULTRA. Moore... Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd generation...systems Viral production Frequently asked questions about lentivirus use Glossary of commonly used lentiviral... or shRNA. See article for detailed information about cloning & additional plasmids. Aebischer and Trono...

... and breathing, and often results in death within 3-5 years from symptom onset. Disease mechanisms are...Find out about the plasmid collections available at Addgene that may be useful for neurodegeneration ...Scientists are studying protein aggregation, learning about the blood brain barrier integrity, and using CRISPR...with support from the BRAIN Initiative. Jackson Laboratory (Link opens in a new window) A collection of ...

...variant 3' NGAG SpCas9 VQR variant  3' NGAN or NGNG  xCas9 3' NG, GAA, or GAT SpCas9-NG 3' NG ...SpCas9 3' NGG SpCas9 D1135E variant 3' NGG (reduced NAG binding) SpCas9 VRER variant 3' NGCG...CJ) 3' NNNNRYAC  Neisseria meningitidis (NM) 3' NNNNGATT  Streptococcus thermophilus (ST) 3' NNAGAAW...target sequence contained within the guide RNA and a 3-base pair sequence known as the Protospacer Adjacent...must contain a PAM sequence immediately downstream (3’) of the site targeted by the guide RNA. In the absence...researchers to target many different genomic loci. 3. Cleave target DNA resulting in a double-strand break...mediated DNA damage is a DSB within the target DNA ~3-4 nucleotides upstream of the PAM sequence. Cas9 Species...

...assist with restriction enzyme digestion (usually 3-6bp) Restriction Site: Your chosen restriction site...Primer will use the sequence 5'-ATGTGGCATATCTCGAAGTAC-3' for the region that binds the ORF and we will add...our Forward Primer 5'-GAATTCATGTGGCATATCTCGAAGTAC-3'. Many restriction enzymes do not cut DNA efficiently...more information). Thus, we recommend that you add 3-6 bases upstream of your restriction site to improve... sequence of 5'-TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but...including the stop codon (5'-TGGCATATCTCGAAGTACTGA-3'), then adding NotI (GCGGCCGC) and then TAAGCA to ...sequence of 5'-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3' (30bp with 18bp of homology to the ORF). We now need...

...polymerase leaves a single deoxyadenosine (A) at the 3' ends of PCR products. The complimentary “T” in the...this enzyme recognizes the DNA sequence 5´-(C/T)CCTT-3' and digests double stranded DNA at this sequence.... is stored as a covalent bond between the cleaved 3′ DNA strand and a tyrosyl residue of topoisomerase... provide vectors or cloning arms with overhanging 3´ deoxythymidine (T) residues that are covalently linked...potential positive clones. Once you introduce your 3’-end “A” overhang insert, the magic of TOPO cloning...Reaction: Mix together the PCR product and TOPO Vector.  3. Incubate 5 Minutes at Room Temperature: You can place...proofreading polymerases will also remove all unpaired 3’ ends in your PCR product. If you need to decrease...

...ligase 25 °C 1 hour 3-base or 4-base T4 DNA ligase 25 °C 18 hours 3-base or 4-base T4 ...ligase 37 °C 1 hour 3-base or 4-base T4 DNA ligase 37 °C 18 hours 3-base or 4-base Bbsl-HF...-16 °C cycling 3-base Sapl, 1x NEBridge Ligase MM 37-16 °C cycling 3-base Table 1: Enzyme... the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same three nucleotides in a...could be done using overhang sets that broke rules 3—5. Fidelity, it turns out, might depend more on the...must be used. Each of the tools works with either 3-base or 4-base overhangs, and with a variety of different...BspQI, 1x NEBridge Ligase MM 42-16 °C cycling 3-base  PaqCl®, 1x T4 DNA Ligase Buffer 37-16 ...

...Figure 3: BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations of IL-3. Cells..., which depend on IL-3 to proliferate. The cells cultured with a low dose of IL-3 showed reduced proliferation...high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...Cells grown in high concentrations of IL-3 show a high proportion of blue fluorescence, indicating cell ...proliferation. BaF3 cells maintained in media without IL-3 show only red fluorescence. Image from Eastman et ...Streptococcus canis Cas9 nuclease that has a broad 5′-NNG-3′ PAM. Get the plasmids at Addgene The Human GlycoGene... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's...

...the PITCh vector should be constructed with 5’ and 3’ microhomology to the insertion locus flanking the...the GFP-Puro cassette and one in between the 5’ and 3’ microhomologies in the genomic locus. The first two...breaks allow for two sets of microhomologies (5’ and 3’) to anneal, knocking the GFP-Puro cassette into the...plasmid contains a GFP-Puro cassette flanked by 5' and 3' microhomology and PITCh-gRNA binding sites. This ...microhomologies in the PITCh vector ~20 bp 5’ and 3’ microhomologies are added to the GFP-Puro cassette...a vector containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector carrying...of clones displayed proper insertion at the 5’ and 3’ junctions, respectively. These results indicate that...

...bigger than ∼3 kb are packaged less efficiently. Subcomponents: U3; Unique 3'; region at the 3' end of viral...SIN γ-retrovirus lack most of the U3 region of the 3’ LTR and are primarily used in gene therapy research...viral genomic RNA (but found at both the 5' and 3' ends of the provirus). Contains sequences necessary...transcription. R; Repeat region found within both the 5' and 3' LTRs of retro/lentiviral plasmids. U5; Unique 5';...the viral genomic RNA (but found at both the 5' and 3' ends of the provirus). 5’ LTR Acts as an RNA...proceeds through U5 and the rest of the provirus. 3’ LTR Terminates transcription started by 5' LTR ...packaging methods. The three components described above (envelope, packaging, and transfer) are supplied...

...genetic tools facilitates plant science. Challenge #3: Intellectual property There is another challenge ...Revolution Bioengineering. Read their previous blog post about how they started their company here. The first transgenic... foundation of life on earth, and opportunities abound to build better fuels, feeds, foods, and fibers... we hope to inspire and open a new conversation about GMOs.  Challenge #2: Technical obstacles to plant...not understand, and the complicated and unique metabolic processes of plants are being investigated on ...biochemical pathways involved in central anthocyanin metabolism are well understood. Additional work has also...knockouts to better understand developmental, metabolic, and photosynthetic processes. Often the tools...

...contains a potential cleavage site for furin proteases.( 3 ) BSG - (CD147), transmembrane glycoprotein of the...lightning speed to elucidate as much as they can about this novel virus. It is now known that SARS-CoV-...and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner, et al. bioRxiv 2020.06.23.166397...

...market; you need one with 5’->3’ polymerase activity (for amplification), 3’->5’ exonuclease activity (increases...on either side of the desired mutation (usually 1-3 mismatched bases) is sufficient for your primers to...present, a minor extension can usually ensure that the 3’-base(s) do not form secondary structures. The introduction...template (e.g. 0.1-1.0 ng/μl). Smaller plasmids (~3 kb) are generally more efficiently amplified than ...addition of DMSO to the PCR reaction (usually around 3% final concentration). The DMSO reduces secondary ...(increases amplification fidelity), and no 5’->3’ exonuclease activity (which could potentially truncate... TA-cloning). Such non-complementary bases at the 3’-end interfere with plasmid reconstitution, and consequently...

... (Nishimasu et al., 2014). Figure 3: pegRNAs whose 3′ extension begins with a C base can cause...Created with BioRender.com. The first base of the 3′ extension of the pegRNA should not be C. A C base...Add silent mutations near point mutations to create 3-base (or longer) tracts of edited bases. DNA mismatch...targets single-base mismatches (left), but “bubbles” of 3 or more mismatched bases can often evade MMR (right...epegRNAs: epegRNAs include structured motifs at their 3′ end to protect them from degradation. When designing...protein La to protect the end of the pegRNA. Adding 3′ polyU tracts to the end of pegRNAs (but not epegRNAs...prime editing efficiency. Nature Biotechnology, 40(3), 402–410. https://doi.org/10.1038/s41587-021-01039...

...overlap between them (Table 1). In each construct, 3 copies of the fluorescent protein were fused in-frame...from the tyrosine-protein kinase Lyn) Membranes 3 mCherry 587nm/ 610 nm human β-Actin Actin 4 Citrine...sites of the MCS are only regenerated at the 5’ and 3’ termini of the ligation product, and the restriction... them, but rather, can only be added to the 5’ or 3’ terminus using the newly generated MCS. MXS-chaining...the third module is added to the 5’ terminus or the 3’ terminus of the ligation product. Comparison to other...chaining-based cloning methods are listed in Table 3. The goal of high throughput cloning is to facilitate...Chaining by emailing us at [email protected]. Table 3: Chaining Based Cloning Methods Method Restriction...

...hype. 3. “Passion gives you energy.” What is your passion? Even if no one else is excited about it, distinguish...success? We asked leading professors for their top 3 tips. Tom Ellis 1. Keep up with the literature. ... that your experiment is definitely not a fluke.” 3. Engage in various projects and help colleagues with... competitive. Be curious and interdisciplinary.   3. “There are many different ways to succeed – as loner...: Read the Newest Posts in this Blog Series   Our 3 PIs answer more questions, including: Advice for ... passionate about. Check out my previous post if you're looking for more information about the benefits...every spurious result under the sun. Academia is about convincing results, and nothing is more convincing...

...information about the aims and approach of CeNGEN, check out their publication.   3’UTRome The 3’UTRs of ... the role of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a 3’UTR database...This database provides detailed information about C. elegans 3’UTRs structures, alternative polyadenylation...Mangone, Marco, et al. "UTRome. org: a platform for 3′ UTR biology in C. elegans." Nucleic acids research...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...resources on the Addgene blog Learn more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids...always emphasized the need for open science and collaboration. The field already has comprehensive reference...

...: a review of the epidemiology. J Pediatric . 148(3):347-352. (Link opens in a new window) PMID: 16615965... working with the RSRT along with individual laboratories to assemble a Rett Syndrome plasmid resource...contact information by following the link to their laboratory website. Mouse Line Mutation (DNA) Background...contact information by following the link to their laboratory website. Cell Line Mutation (DNA) Mutation (protein...