{"id":2260,"date":"2018-12-04T10:55:10","date_gmt":"2018-12-04T01:55:10","guid":{"rendered":"http:\/\/163.180.4.222\/lab\/?p=2260"},"modified":"2018-12-04T10:55:10","modified_gmt":"2018-12-04T01:55:10","slug":"a-dna-nanomachine-chemically-resolves-lysosomes-in-live-cells","status":"publish","type":"post","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=2260","title":{"rendered":"A DNA nanomachine chemically resolves lysosomes in live cells"},"content":{"rendered":"

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Abstract<\/h5>\n
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Lysosomes are multifunctional, subcellular organelles with roles in plasma membrane repair, autophagy, pathogen degradation and nutrient sensing. Dysfunctional lysosomes underlie Alzheimer\u2019s disease, Parkinson\u2019s disease and rare lysosomal storage diseases, but their contributions to these pathophysiologies are unclear. Live imaging has revealed lysosome subpopulations with different physical characteristics including dynamics, morphology or cellular localization. Here, we chemically resolve lysosome subpopulations using a DNA-based combination reporter that quantitatively images pH and chloride simultaneously in the same lysosome while retaining single-lysosome information in live cells. We call this technology two-ion measurement or 2-IM. 2-IM of lysosomes in primary skin fibroblasts derived from healthy individuals shows two main lysosome populations, one of which is absent in primary cells derived from patients with Niemann\u2013Pick disease. When patient cells are treated with relevant therapeutics, the second population re-emerges. Chemically resolving lysosomes by 2-IM could enable decoding the mechanistic underpinnings of lysosomal diseases, monitoring disease progression or evaluating therapeutic efficacy.<\/p>\n<\/div>\n<\/div>\n<\/section>\n

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Fig. 1: Design and characterization of ChloropHore.<\/b><\/figcaption>
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a<\/b>, Schematic of the working principle. A pH-induced change in fluorescence resonance energy transfer between Alexa\u2009546 (donor, orange sphere) and Alexa\u2009647 (acceptor, red star) reports pH ratiometrically. A Cl\u2212<\/sup>-sensitive fluorophore (BAC, green triangle) and Alexa\u2009647 reports Cl\u2212<\/sup>\u00a0ratiometrically. P, Cl\u2212<\/sup>-sensing module (purple line) containing a Cl\u2212<\/sup>-sensitive fluorophore BAC; C1, pH-sensing module containing i-motif sequence (grey line); C2, normalizing module (orange line) containing a pH, Cl\u2212<\/sup>-insensitive fluorophore Alexa 647.\u00a0b<\/b>, pH and Cl\u2212<\/sup>\u00a0response profiles of ChloropHore. Normalized fluorescence intensity ratio (D\/A) of donor (D) and acceptor (A) on donor excitation in\u2009vitro as a function of pH and 50\u2009mM Cl\u2212<\/sup>(red). Normalized fluorescence intensity ratio (R\/G) of Alexa\u2009647 (R) and BAC (G) as a function of Cl\u2212<\/sup>\u00a0concentration at pH\u20097 (blue). Values were normalized to R\/G at 5\u2009mM Cl\u2212<\/sup>\u00a0or D\/A at pH 4.\u00a0c<\/b>, Performance of the pH-sensing module (red) at different [Cl\u2212<\/sup>] and of the Cl\u2212<\/sup>-sensing module at different pH values (blue). Fold changes in D\/A (red bars) or R\/G (dark blue bars) for the pH- and Cl\u2212<\/sup>-sensing modules are shown. Stern\u2013Volmer constant (K<\/i>SV<\/sub>, light blue bars) for Cl\u2212<\/sup>\u00a0sensing at each pH is also shown.\u00a0d<\/b>,e<\/b>, Calibration surface plots of D\/A (d<\/b>) and R\/G (e<\/b>) of ChloropHore as a function of Cl\u2212<\/sup>\u00a0and pH. Error bars indicate the mean\u2009\u00b1\u2009standard error of the mean (s.e.m.) of three independent measurements.<\/p>\n<\/div>\n<\/div>\n

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\uc6d0\ubb38: \uc5ec\uae30<\/a>\ub97c \ud074\ub9ad\ud558\uc138\uc694~<\/p>\n

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    Abstract Lysosomes are multifunctional, subcellular organelles with roles in plasma membrane repair, autophagy, pathogen degradation and nutrient sensing. Dysfunctional lysosomes underlie Alzheimer\u2019s disease,(more…)<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"jetpack_post_was_ever_published":false,"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":true,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2}},"categories":[33,34,29,30],"tags":[],"class_list":["post-2260","post","type-post","status-publish","format-standard","hentry","category-do-biology","category-lets-do-chemistry","category-lets-do-science","category-recent-science-news"],"aioseo_notices":[],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"","jetpack-related-posts":[{"id":4699,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=4699","url_meta":{"origin":2260,"position":0},"title":"Senescent cells feed on their neighbours","author":"biochemistry","date":"October 31, 2019","format":false,"excerpt":"\u00a0 Chemotherapy-treated cancer cells that enter a non-dividing state called senescence can nevertheless boost cancer growth. The finding that these cells eat neighbouring cells reveals a mechanism that enables senescent cells to persist. \u00a0 \u00a0 Multicellular life requires individual cells to cooperate in a way that benefits the organism. Cells\u2026","rel":"","context":"In "Let's Do Biology!"","block_context":{"text":"Let's Do Biology!","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=33"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]},{"id":3524,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3524","url_meta":{"origin":2260,"position":1},"title":"Stem-cell and genetic therapies make a healthy marriage","author":"biochemistry","date":"May 16, 2019","format":false,"excerpt":"\u00a0 \u00a0 This scientific partnership could fight everything from blood diseases to HIV. \u00a0 A culture from human tissue showing neurons derived from stem cells.Credit: Daniel Schroen\/Cell Applications Inc\/SPL \u00a0 \u00a0 Aside from a 20-second exposure to the outside world at birth, David Vetter spent his entire life cocooned in\u2026","rel":"","context":"In "Essays on Science"","block_context":{"text":"Essays on Science","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=32"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]},{"id":2252,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=2252","url_meta":{"origin":2260,"position":2},"title":"The paradox of mutations and cancer","author":"biochemistry","date":"December 3, 2018","format":false,"excerpt":"\u00a0 \u00a0 The past decade has witnessed the cataloging of genetic mutations in cancer genomes, providing new insights into how and in what ways cancer can develop and spread (1,\u00a02). The focus has been on defining specific \u201cdriver\u201d mutations, genetic errors in cancer cells that reveal basic biological processes gone\u2026","rel":"","context":"In "Essays on Science"","block_context":{"text":"Essays on Science","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=32"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]},{"id":2582,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=2582","url_meta":{"origin":2260,"position":3},"title":"Technologies to watch in 2019","author":"biochemistry","date":"January 29, 2019","format":false,"excerpt":"\u00a0 \u00a0 From higher-resolution imaging to genome-sized DNA molecules built from scratch, the year ahead looks exciting for life-science technology. \u00a0 An automated bioreactor system for growing yeast, which can be used to investigate synthetic genomes \u2014 one area poised to make big strides this year.Credit: Tim Llewellyn\/Ginkgo Bioworks \u00a0\u2026","rel":"","context":"In "Essays on Science"","block_context":{"text":"Essays on Science","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=32"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]},{"id":4854,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=4854","url_meta":{"origin":2260,"position":4},"title":"Got mutation? \u2018Base editors\u2019 fix genomes one nucleotide at a time","author":"biochemistry","date":"November 19, 2019","format":false,"excerpt":"\u00a0 \u00a0 A new class of CRISPR-based tools efficiently corrects point mutations in cell lines, animal models and perhaps the clinic. \u00a0 \u00a0 Credit: Getty \u00a0 \u00a0 When Xingxu Huang began thinking about correcting disease-causing mutations in the human genome, his attention turned naturally to CRISPR\u2013Cas9. But it quickly became\u2026","rel":"","context":"In "Let's Do Biology!"","block_context":{"text":"Let's Do Biology!","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=33"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]},{"id":3893,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3893","url_meta":{"origin":2260,"position":5},"title":"Epigenetics comes to RNA","author":"biochemistry","date":"July 16, 2019","format":false,"excerpt":"\u00a0 \u00a0 An enzyme (pink) places a chemical mark (gold) on messenger RNA (blue), in an artist's concept. IMAGE: STORM THERAPEUTICS \u00a0 \u00a0 The idea that chemical tags on genes can affect their expression without altering the DNA sequence, once surprising, is the stuff of textbooks. The phenomenon, epigenetics, has\u2026","rel":"","context":"In "Let's Do Biology!"","block_context":{"text":"Let's Do Biology!","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=33"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]}],"jetpack_sharing_enabled":false,"jetpack_shortlink":"https:\/\/wp.me\/p9Xo1j-As","_links":{"self":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/2260","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=2260"}],"version-history":[{"count":1,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/2260\/revisions"}],"predecessor-version":[{"id":2261,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/2260\/revisions\/2261"}],"wp:attachment":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=2260"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=2260"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=2260"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}