{"id":4118,"date":"2019-09-23T18:46:30","date_gmt":"2019-09-23T09:46:30","guid":{"rendered":"http:\/\/163.180.4.222\/lab\/?p=4118"},"modified":"2019-09-23T18:46:30","modified_gmt":"2019-09-23T09:46:30","slug":"crispr-mediated-live-imaging-of-genome-editing-and-transcription","status":"publish","type":"post","link":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=4118","title":{"rendered":"CRISPR-mediated live imaging of genome editing and transcription"},"content":{"rendered":"

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Tracking nucleic acids in living cells<\/h4>\n

Fluorescence in situ hybridization (FISH) is a powerful molecular technique for detecting nucleic acids in cells. However, it requires cell fixation and denaturation. Wang\u00a0et al.<\/em>\u00a0found that CRISPR-Cas9 protects guide RNAs from degradation in cells only when bound to target DNA. Taking advantage of this target-dependent stability switch, they developed a labeling technique, named CRISPR LiveFISH, to detect DNA and RNA using fluorophore-conjugated guide RNAs with Cas9 and Cas13, respectively. CRISPR LiveFISH improves the signal-to-noise ratio, is compatible with living cells, and allows tracking real-time dynamics of genome editing, chromosome translocation, and transcription.<\/p>\n

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Science<\/em>, this issue p.\u00a01301<\/a><\/p>\n<\/div>\n

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Abstract<\/h4>\n

We report a robust, versatile approach called CRISPR live-cell fluorescent in situ hybridization (LiveFISH) using fluorescent oligonucleotides for genome tracking in a broad range of cell types, including primary cells. An intrinsic stability switch of CRISPR guide RNAs enables LiveFISH to accurately detect chromosomal disorders such as Patau syndrome in prenatal amniotic fluid cells and track multiple loci in human T lymphocytes. In addition, LiveFISH tracks the real-time movement of DNA double-strand breaks induced by CRISPR-Cas9\u2013mediated editing and consequent chromosome translocations. Finally, by combining Cas9 and Cas13 systems, LiveFISH allows for simultaneous visualization of genomic DNA and RNA transcripts in living cells. The LiveFISH approach enables real-time live imaging of DNA and RNA during genome editing, transcription, and rearrangements in single cells.<\/p>\n<\/div>\n

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

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    Tracking nucleic acids in living cells Fluorescence in situ hybridization (FISH) is a powerful molecular technique for detecting nucleic acids in cells. However,(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-4118","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":2672,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=2672","url_meta":{"origin":4118,"position":0},"title":"CRISPR-Cas9-Based Genome Editing of Human Cells","author":"biochemistry","date":"February 15, 2019","format":false,"excerpt":"\u00a0 \u00a0 CRISPR\/Cas9 systems are engineered versions of the Cas9 protein and guide RNA. \u00a0Typically, they are identical to the\u00a0Streptococcus pyogenes\u00a0type II CRISPR systems, except that a single guide-RNA is used in place of the complementary crRNAs and tracrRNAs of the natural CRISPR system, and the Cas9 protein is codon-optimized\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":"Genome Editing Overview2","src":"https:\/\/i0.wp.com\/sites.tufts.edu\/crispr\/files\/2014\/11\/Genome-Editing-Overview2-1024x667.png?resize=350%2C200&ssl=1","width":350,"height":200,"srcset":"https:\/\/i0.wp.com\/sites.tufts.edu\/crispr\/files\/2014\/11\/Genome-Editing-Overview2-1024x667.png?resize=350%2C200&ssl=1 1x, https:\/\/i0.wp.com\/sites.tufts.edu\/crispr\/files\/2014\/11\/Genome-Editing-Overview2-1024x667.png?resize=525%2C300&ssl=1 1.5x, https:\/\/i0.wp.com\/sites.tufts.edu\/crispr\/files\/2014\/11\/Genome-Editing-Overview2-1024x667.png?resize=700%2C400&ssl=1 2x"},"classes":[]},{"id":1857,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=1857","url_meta":{"origin":4118,"position":1},"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 "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":3361,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3361","url_meta":{"origin":4118,"position":2},"title":"When genome editing goes off-target","author":"biochemistry","date":"April 19, 2019","format":false,"excerpt":"\u00a0 \u00a0 Editing DNA in eukaryotic cells with CRISPR-based systems has revolutionized the genome engineering field. Cas (CRISPR-associated) endonucleases are directed to a particular location in the genome by a short guide RNA, providing an easily programmable strategy to target any section of DNA. As of now, two CRISPR-based approaches\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":3891,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=3891","url_meta":{"origin":4118,"position":3},"title":"Inserting DNA with CRISPR","author":"biochemistry","date":"July 16, 2019","format":false,"excerpt":"\u00a0 \u00a0 Most prokaryotes rely on the CRISPR-Cas system for adaptive immunity against viruses and mobile elements (1,\u00a02). Small RNAs produced from CRISPR direct Cas effector proteins to seek and destroy nucleic acids from invaders that have complementary target sites (3). There are multiple types of CRISPR, which are defined\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":4854,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=4854","url_meta":{"origin":4118,"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":4845,"url":"https:\/\/biochemistry.khu.ac.kr\/lab\/?p=4845","url_meta":{"origin":4118,"position":5},"title":"CRISPR tool modifies genes precisely by copying RNA into the genome & CRISPR: the movie","author":"biochemistry","date":"November 15, 2019","format":false,"excerpt":"\u00a0 \u00a0 The ultimate goal of genome editing is to be able to make any specific change to the blueprint of life. A \u2018search-and-replace\u2019 method for genome editing takes us a giant leap closer to this ambitious goal. \u00a0 \u00a0 Variation in the DNA sequences that constitute the blueprint of\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-14q","_links":{"self":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/4118","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=4118"}],"version-history":[{"count":1,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/4118\/revisions"}],"predecessor-version":[{"id":4119,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=\/wp\/v2\/posts\/4118\/revisions\/4119"}],"wp:attachment":[{"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=4118"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=4118"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biochemistry.khu.ac.kr\/lab\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=4118"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}