Dow: >95% of clones had biallelic loss. Could multiplex, have gone up to six guide RNAs.'12 ref https://t.co/QHI6BH81pa #Tricon
7:22pm February 22nd 2017 via Hootsuite
Dow: Induceable Cas9 w/GFP; can target APC and APC/p53. Shows expression loss and in vivo loss of protein #Tricon
7:21pm February 22nd 2017 via Hootsuite
Dow: Many components altered, extra cellular down to nuclear effect. HDR and NHEJ; they use NHEJ for large translocation engineering #Tricon
7:20pm February 22nd 2017 via Hootsuite
Dow: Want to better model human biology. APC drivent by Wnt ligand; bind receptors, beta-catenin stabilized, drives c-Myc others #Tricon
7:18pm February 22nd 2017 via Hootsuite
Dow:CR cancer: 1.4M new cases, 700K deaths ww; 85% carry APC ts. Models do not recapitulate human CRC biology #Tricon
7:17pm February 22nd 2017 via Hootsuite
Lukas Dow (Weill Cornell NY) Tailored pre-clinical models with CRISPR-based genome editing #Tricon
7:16pm February 22nd 2017 via Hootsuite
Q: G12C not scoring highly; connection to particular ts? Winslow: P53del, RASdel data supports; lung ca is smoking-driven thus G12C #Tricon
7:15pm February 22nd 2017 via Hootsuite
Winslow: Have used models for 10y but genotype-spec biology w/phenotype, drug response, lots of time and resources. CRISPR efficient #Tricon
7:10pm February 22nd 2017 via Hootsuite
Winslow: Tumors induced by HDR in KRAS, shows cancer cell #'s of G1R, G12D, G12R over logs of cell numbers #Tricon
7:05pm February 22nd 2017 via Hootsuite
Winslow: AAV vector delivered; 4-10 months, showed lung imaging and H&E stains. Looked across 50 tumors; showed G12D/V/R prev #Tricon
7:04pm February 22nd 2017 via Hootsuite
Winslow: Created a panel of KRAS codon 12/13 mutations, in equal proportions, see % of tumors formed. #Tricon
7:03pm February 22nd 2017 via Hootsuite
Winslow: CRISPR causes ds breaks; can do HDR with mutation. Created template in AAV HDR template to repair RAS allele #Tricon
7:02pm February 22nd 2017 via Hootsuite
Winslow: Oncogenic mutations - highlights KRAS, 'all studies use G12D', but they may have different effects on phenotype, drug resp #Tricon
7:01pm February 22nd 2017 via Hootsuite
Winslow: There are many coincident tumor suppressor mutations. P53 for example; distibution w/LKB1mut is different #Tricon
6:58pm February 22nd 2017 via Hootsuite
Winslow: Shows data - relative size as a percentile; Lkb1 had the highest effect Setd2 next one (chr regulator) #Tricon
6:56pm February 22nd 2017 via Hootsuite
Winslow: Can look at the effect of deleting all these ts genes. Shows lung lobe with a lot of tumors; H&E stain also shown #Tricon
6:55pm February 22nd 2017 via Hootsuite
Winslow: Took 11 tumor suppressors, each guide RNA has an ID, and each tumor has a barcode. Reading genotype and size #Tricon
6:54pm February 22nd 2017 via Hootsuite
Winslow: The whole lung, grind it up, and quantify each cell # from a bulk tumor. #Tricon
6:53pm February 22nd 2017 via Hootsuite
Winslow: Use lentiviral vectors; stable integration; library w/1M barcodes, so each infected cell is labeled. #Tricon
Winslow: Put diverse genotypes into one mouse; replacing floxed alleles with CRISPR 'a few thousand tumors' #Tricon
6:52pm February 22nd 2017 via Hootsuite
Winslow: Many tumor suppressors - long list from TCGA in lung adenocarcinomas. Able to inactivate genes, lentiviral sgRNAs #Tricon
6:51pm February 22nd 2017 via Hootsuite
Winslow: Also poor quantification of tumor response (in vivo imaging, histology) #Tricon
6:49pm February 22nd 2017 via Hootsuite
Winslow: Major limitation for Cre/Lox engineered mouse models for translation - limited diversity; diificult to scale-up #Tricon
Monte Winslow (Stanford Univ CA) Cancer modeling with in vivo CRISPR/Cas9 genome editing #Tricon
6:47pm February 22nd 2017 via Hootsuite
Smith: Size-based separation shown of exosomes. Can do viruses but have not done that work yet. #Tricon
6:26pm February 22nd 2017 via Hootsuite
Smith: docetaxel-resistant prostate ca-derived exosomes (collab with Mt Sinai) found to be smaller than sensitive ones #Tricon
6:25pm February 22nd 2017 via Hootsuite
Smith: Air-driven pump, for the fluid to push it through. Finishes with ability to subfractionate the exosome population #Tricon
6:24pm February 22nd 2017 via Hootsuite
Smith: How to use it, a way to get fluid into the chip itself? Decoupling via chip into a cartridge, up to 1mL input #Tricon
6:23pm February 22nd 2017 via Hootsuite
Smith: Covers technology for extracting sample from the chip. 50-100uL/hr per layer with stackable ability #Tricon
6:22pm February 22nd 2017 via Hootsuite
Smith: Dealing with clogging: use filtration before loading at 0.2um. Developed sorting pillar array, to run to full clog w/beads #Tricon
6:20pm February 22nd 2017 via Hootsuite
Smith:Thus they can create a device to separate on size but overcome dilution problems. Full-width injection. #Tricon
6:16pm February 22nd 2017 via Hootsuite
Smith: Posts can be changed in ordering, diameter. Simulated model w/predictions, verify experimentally. Showed micrographs, data #Tricon
6:15pm February 22nd 2017 via Hootsuite
Smith: Integrated the Gaussian distribution curves. Can get almost perfect separation of 200bp difference. #Tricon
6:14pm February 22nd 2017 via Hootsuite
Smith: Can do multimodal - shows clean separation of the 100bp, to 1kb, to 10kb. Borrowed chromatography resolution curves #Tricon
6:12pm February 22nd 2017 via Hootsuite
Smith: Shows at 10kb all aligned on the side of the channel. Thus opportunities for DNA seq prep. #Tricon
6:11pm February 22nd 2017 via Hootsuite
Smith: Shows cool movies - 100bp, 1kb, and 10kb DNAs in the inlet and the outlet. Gap size the same; shows zig-zag 100bp #Tricon
Smith: Biomarker separation down to 20ng - first beads, then exosomes, then 10kb from 100kb DNAs #Tricon
6:09pm February 22nd 2017 via Hootsuite
Smith: Can subfractionate, get compartimentalization. https://t.co/Y5lSTSBjbj Shows '16 ref https://t.co/BLdxE4urM2 #Tricon
6:08pm February 22nd 2017 via Hootsuite
Smith: Exosomes are 30-150nm; DNA is 2nm; viruses is 20-200nm. Streamlines among pillars: larger particles go to assymetry #Tricon
6:05pm February 22nd 2017 via Hootsuite
Smith: DLD is Deterministic Lateral Displacement. It is continuous flow, can separate different sized molecules, simple design #Tricon
6:04pm February 22nd 2017 via Hootsuite
Smith: Workflow for manual extraction for ctDNA shown. Microfluidics have many papers, many techniques and approaches #Tricon
6:03pm February 22nd 2017 via Hootsuite
Smith: Shows 1-2h Ultracent at 10k xg; then 2-3h at 100k xg, then 15h sucrose gradient, then anti-CD63 2-3h #Tricon
6:02pm February 22nd 2017 via Hootsuite
Smith: Shows figure from '15 rev https://t.co/V8aBglOBTC on importance of exosomes and involvement with disease #Tricon
6:01pm February 22nd 2017 via Hootsuite
Smith: Nanofabrication strategy includes flexible electronics, quantum computing, IoT, neuromorphic computing, nanobiotech #Tricon
5:59pm February 22nd 2017 via Hootsuite
Smith: IBM Research synonymous with Watson. nanoDLD on-chip separation of exosomes and nucleic acids. #Tricon
5:58pm February 22nd 2017 via Hootsuite
Joshua Smith (IBM Nanobiotechnology Group NY): Sample prep for liquid biopsies on a chip: exosomes, DNA and beyond #Tricon
5:57pm February 22nd 2017 via Hootsuite
Q: Does read-until affect nanopore life? A: ONT says it doesn't affect the pore. Sandia can't confirm - sees some variability. #Tricon
5:54pm February 22nd 2017 via Hootsuite
Q: How to deal with tubing contamination? Patel: Use disposable tubing. #Tricon
5:53pm February 22nd 2017 via Hootsuite
Q: Miniaturizing involves smaller volumes? Patel: Volumes are on the order of 10-100uL just making equipment smaller #Tricon
5:47pm February 22nd 2017 via Hootsuite
Patel: Library prep takes 3h manually. #Tricon
5:46pm February 22nd 2017 via Hootsuite
