{"id":3261,"date":"2019-04-09T15:49:35","date_gmt":"2019-04-09T06:49:35","guid":{"rendered":"http:\/\/163.180.4.222\/lab\/?p=3261"},"modified":"2019-04-09T15:49:35","modified_gmt":"2019-04-09T06:49:35","slug":"highly-efficient-expression-of-circular-rna-aptamers-in-cells-using-autocatalytic-transcripts","status":"publish","type":"post","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3261","title":{"rendered":"Highly efficient expression of circular RNA aptamers in cells using autocatalytic transcripts"},"content":{"rendered":"<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p id=\"Abs1\" class=\"js-section-title section-title strong position-relative tighten-line-height background-gray-light pt20 pb6 pl0 pr20 standard-space-below small-space-above mq640-pt10 mq640-pb10 mq640-pl20 mq640-mt0 mq640-ml-20 mq640-mr-20 extend-left\"><strong>Abstract<\/strong><\/p>\n<div id=\"Abs1-content\" class=\"pl20 mq875-pl0 js-collapsible-section\">\n<p>&nbsp;<\/p>\n<p>RNA aptamers and RNA aptamer-based devices can be genetically encoded and expressed in cells to probe and manipulate cellular function. However, their usefulness in the mammalian cell is limited by low expression and rapid degradation. Here we describe the Tornado (Twister-optimized RNA for durable overexpression) expression system for achieving rapid RNA circularization, resulting in RNA aptamers with high stability and expression levels. Tornado-expressed transcripts contain an RNA of interest flanked by Twister ribozymes. The ribozymes rapidly undergo autocatalytic cleavage, leaving termini that are ligated by the ubiquitous endogenous RNA ligase RtcB. Using this approach, protein-binding aptamers that otherwise have minimal effects in cells become potent inhibitors of cellular signaling. Additionally, an RNA-based fluorescent metabolite biosensor for\u00a0<i>S<\/i>-adenosyl methionine (SAM) that is expressed at low levels when expressed as a linear RNA achieves levels sufficient for detection of intracellular SAM dynamics when expressed as a circular RNA. The Tornado expression system thus markedly enhances the utility of RNA-based approaches in the mammalian cell.<\/p>\n<\/div>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>(\uc6d0\ubb38: <a href=\"https:\/\/www.nature.com\/articles\/s41587-019-0090-6?utm_source=feedburner&amp;utm_medium=feed&amp;utm_campaign=Feed%3A+nbt%2Frss%2Fcurrent+%28Nature+Biotechnology+-+Issue%29\">\uc5ec\uae30<\/a>\ub97c \ud074\ub9ad\ud558\uc138\uc694~)<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>&nbsp; &nbsp; Abstract &nbsp; RNA aptamers and RNA aptamer-based devices can be genetically encoded and expressed in cells to probe and manipulate cellular function. However,<a href=\"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3261\" class=\"more-link\">(more&#8230;)<\/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-3261","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":3304,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3304","url_meta":{"origin":3261,"position":0},"title":"Structure and functional reselection of the Mango-III fluorogenic RNA aptamer","author":"biochemistry","date":"April 16, 2019","format":false,"excerpt":"\u00a0 \u00a0 Abstract \u00a0 Several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal\u2026","rel":"","context":"In &quot;Let's Do Chemistry!&quot;","block_context":{"text":"Let's Do Chemistry!","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=34"},"img":{"alt_text":"","src":"","width":0,"height":0},"classes":[]},{"id":4481,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=4481","url_meta":{"origin":3261,"position":1},"title":"RNA therapies explained","author":"biochemistry","date":"October 18, 2019","format":false,"excerpt":"\u00a0 Treatments that target RNA or deliver it to cells fall into three broad categories, with hybrid approaches also emerging. \u00a0 \u00a0 Illustration of messenger RNA (red) produced from a DNA strand (purple).\u00a0Credit: Juan Gaertner\/SPL \u00a0 \u00a0 The conventional view of RNA casts the molecule in a supporting role \u2014\u2026","rel":"","context":"In &quot;Let's Do Biology!&quot;","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":2099,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=2099","url_meta":{"origin":3261,"position":2},"title":"Aptamers and Clinical Applications","author":"biochemistry","date":"October 23, 2018","format":false,"excerpt":"\u00a0 \u00a0 Gov Lists 28 Clinical Studies, Mostly Ocular, for Aptamers Only 3 On This List Are Currently Active Studies NOXXON Pharma\u2019s Mirror-Image L-RNA Aptamer is in the Clinic for Cancers \u00a0 Devoted readers of\u00a0Zone In With Zon\u00a0who have photographic memories\u2014or anyone who simply uses this blog\u2019s search engine\u2014will know\u2026","rel":"","context":"In &quot;Essays on Science&quot;","block_context":{"text":"Essays on Science","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?cat=32"},"img":{"alt_text":"","src":"https:\/\/i0.wp.com\/zon.trilinkbiotech.com\/wp-content\/uploads\/2018\/02\/aptamer-300x242.png?resize=350%2C200","width":350,"height":200},"classes":[]},{"id":3448,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3448","url_meta":{"origin":3261,"position":3},"title":"Pinpointing a spatial address for RNA profiles in tissues","author":"biochemistry","date":"May 4, 2019","format":false,"excerpt":"\u00a0 \u00a0 Knowing the gene-expression pattern of individual cells can unlock their identity. A refined method for generating cellular RNA profiles offers a way to obtain such data at a high level of spatial resolution in intact tissues. \u00a0 \u00a0 Monitoring messenger RNA in cells is a way to gather\u2026","rel":"","context":"In &quot;Essays on Science&quot;","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":1316,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=1316","url_meta":{"origin":3261,"position":4},"title":"Technique to measure the expression dynamics of each gene in a single cell","author":"biochemistry","date":"August 9, 2018","format":false,"excerpt":"\u00a0 \u00a0 (\uc6d0\ubb38) \u00a0 \u00a0 A method has been developed to infer whether the expression of each gene in a single cell is increasing or decreasing, and at what rate, using RNA-sequencing data. This tool has many potential applications. \u00a0 \u00a0 To understand and control complex systems, we must be\u2026","rel":"","context":"In &quot;Let's Do Biology!&quot;","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":1857,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=1857","url_meta":{"origin":3261,"position":5},"title":"CRISPR-Cas9 nuclease \uad00\ub828 \uba87 \uac00\uc9c0 \ub274\uc2a4","author":"biochemistry","date":"September 25, 2018","format":false,"excerpt":"\u00a0 \u00a0 CRISPR-Cas9\uacfc \uad00\ub828\ub41c \uba87 \uac00\uc9c0 \uc18c\uc2dd\uc785\ub2c8\ub2e4. (\uc6d0\ubb38: \uc5ec\uae30\ub97c \ud074\ub9ad\ud558\uc138\uc694~) \u00a0 CRISPR tool puts RNA on the record \u00a0 The bacterial-defence system CRISPR\u2013Cas can store DNA snippets that correspond to encountered viral RNA sequences. One such system has now been harnessed to record gene expression over time in bacteria. \u00a0\u2026","rel":"","context":"In &quot;Let's Do Biology!&quot;","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-QB","_links":{"self":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/3261","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=3261"}],"version-history":[{"count":1,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/3261\/revisions"}],"predecessor-version":[{"id":3262,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/3261\/revisions\/3262"}],"wp:attachment":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=3261"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=3261"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=3261"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}