{"id":36,"date":"2021-02-24T14:06:56","date_gmt":"2021-02-24T14:06:56","guid":{"rendered":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/?page_id=36"},"modified":"2021-02-24T15:12:01","modified_gmt":"2021-02-24T15:12:01","slug":"crispr-based-genetic-screens-in-mammalian-cells","status":"publish","type":"page","link":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/research\/crispr-based-genetic-screens-in-mammalian-cells\/","title":{"rendered":"CRISPR-based Genetic Screens in Mammalian Cells"},"content":{"rendered":"<h2>Background<\/h2>\n<p>CRISPRs (clustered regularly interspaced short palindromic repeats) are a family of short DNA sequences present in prokaryotes.  The CRISPR sequences in prokaryotes are originally derived from infectious phages, and help guide the enzyme Cas9 endonuclease to cleave cognate viral DNA sequences as part of a prokaryotic anti-viral response system.  Since 2013, CRISPR-Cas9 technology has been harnessed to perform genome editing as well as targeted gene activation and interference in mammalian cells.<\/p>\n<p>Our laboratory uses CRISPR knockout, activation, and interference libraries in mammalian cells (including induced pluripotent stem cells) to determine the genetic pathways responsible for a variety of fundamental processes, including regulation of <a href=\"https:\/\/www.nature.com\/articles\/s41420-018-0135-5\" target=\"_blank\">cell death<\/a>, <a href=\"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/mBio-2020-Chidawanyika.pdf\" rel=\"noopener\" target=\"_blank\">oxidative stress<\/a>, cell size, organelle mass, and the turnover and trafficking of specific proteins.<\/p>\n<h2>Images<\/h2>\n<div id='gallery-1' class='gallery galleryid-36 gallery-columns-2 gallery-size-large'><figure class='gallery-item'>\n\t\t\t<div class='gallery-icon landscape'>\n\t\t\t\t<a href='https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic.jpg'><img loading=\"lazy\" decoding=\"async\" width=\"740\" height=\"496\" src=\"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-1024x687.jpg\" class=\"attachment-large size-large\" alt=\"Schematic diagram of a CRISPR-based genetic screen in a library of cultured mammalian cells. Results are obtained by (NGS) next-generation sequencing, and analyzed bioinformatically.\" aria-describedby=\"gallery-1-106\" srcset=\"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-1024x687.jpg 1024w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-300x201.jpg 300w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-768x515.jpg 768w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-82x55.jpg 82w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-1600x1074.jpg 1600w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-800x537.jpg 800w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/CRISPR-schematic-580x389.jpg 580w\" sizes=\"auto, (max-width: 740px) 100vw, 740px\" \/><\/a>\n\t\t\t<\/div>\n\t\t\t\t<figcaption class='wp-caption-text gallery-caption' id='gallery-1-106'>\n\t\t\t\tSchematic diagram of a CRISPR-based genetic screen in a library of cultured mammalian cells.  Results are obtained by (NGS) next-generation sequencing, and analyzed bioinformatically.\n\t\t\t\t<\/figcaption><\/figure><figure class='gallery-item'>\n\t\t\t<div class='gallery-icon landscape'>\n\t\t\t\t<a href='https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies.jpg'><img loading=\"lazy\" decoding=\"async\" width=\"740\" height=\"567\" src=\"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-1024x784.jpg\" class=\"attachment-large size-large\" alt=\"Microscopic image of induced Pluripotent Stem Cells (iPSCs) embryoid bodies grown on Matrigel.\" aria-describedby=\"gallery-1-108\" srcset=\"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-1024x784.jpg 1024w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-300x230.jpg 300w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-768x588.jpg 768w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-72x55.jpg 72w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-1600x1225.jpg 1600w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-800x613.jpg 800w, https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-content\/uploads\/sites\/80\/2021\/02\/Embryoid-bodies-580x444.jpg 580w\" sizes=\"auto, (max-width: 740px) 100vw, 740px\" \/><\/a>\n\t\t\t<\/div>\n\t\t\t\t<figcaption class='wp-caption-text gallery-caption' id='gallery-1-108'>\n\t\t\t\tMicroscopic image of induced Pluripotent Stem Cells (iPSCs) embryoid bodies grown on Matrigel.\n\t\t\t\t<\/figcaption><\/figure>\n\t\t<\/div>\n\n","protected":false},"excerpt":{"rendered":"<p>Background CRISPRs (clustered regularly interspaced short palindromic repeats) are a family of short DNA sequences present in prokaryotes. The CRISPR sequences in prokaryotes are originally derived from infectious phages, and help guide the enzyme Cas9 endonuclease to cleave cognate viral DNA sequences as part of a prokaryotic anti-viral response system. [\u2026] <\/p>\n<div class=\"clear\"><\/div>\n<p><a class=\"more_link clearfix\" href=\"https:\/\/geiselmed.dartmouth.edu\/supattapone\/research\/crispr-based-genetic-screens-in-mammalian-cells\/\" rel=\"nofollow\">Read More<\/a><\/p>\n","protected":false},"author":2,"featured_media":0,"parent":32,"menu_order":2,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-36","page","type-page","status-publish","hentry","author-2"],"_links":{"self":[{"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/pages\/36","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/comments?post=36"}],"version-history":[{"count":13,"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/pages\/36\/revisions"}],"predecessor-version":[{"id":119,"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/pages\/36\/revisions\/119"}],"up":[{"embeddable":true,"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/pages\/32"}],"wp:attachment":[{"href":"https:\/\/geiselmed.dartmouth.edu\/supattapone\/wp-json\/wp\/v2\/media?parent=36"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}