{"id":2413,"date":"2020-08-23T09:52:00","date_gmt":"2020-08-23T09:52:00","guid":{"rendered":"https:\/\/dnaera.com\/ro\/?p=2413"},"modified":"2023-04-17T09:22:25","modified_gmt":"2023-04-17T07:22:25","slug":"crispr-cas9-nastroj-na-upravu-dna-oceneny-nobelovou-cenou","status":"publish","type":"post","link":"https:\/\/dnaera.com\/ro\/blog\/crispr-cas9-nastroj-na-upravu-dna-oceneny-nobelovou-cenou\/","title":{"rendered":"CRISPR\/Cas9: n\u00e1stroj na \u00fapravu DNA ocenen\u00fd Nobelovou cenou"},"content":{"rendered":"\n

V\u00f4bec po prv\u00fdkr\u00e1t v hist\u00f3rii boli tento rok ocenen\u00e9 Nobelovou cenou za ch\u00e9miu dve \u017eeny<\/strong>. Americk\u00e1 vedky\u0148a Jennifer Doudna<\/strong>, profesorka na Kalifornskej univerzite v meste Berkeley a franc\u00fazska vedky\u0148a Emmanuelle Charpentier<\/strong>, profesorka, geneti\u010dka, mikrobiologi\u010dka a riadite\u013eka \u00fastavu Maxa Plancka pre infek\u010dn\u00fa biol\u00f3giu.<\/p>\n\n\n\n

Spolo\u010dne objavili met\u00f3du na cielen\u00fa \u00fapravu genetickej inform\u00e1cie (DNA) v rastlinn\u00fdch, \u017eivo\u010d\u00ed\u0161nych, ako aj v \u013eudsk\u00fdch bunk\u00e1ch.<\/strong>\u00a0Hlavn\u00fdm zameran\u00edm v\u00fdskumu oboch vedk\u00fd\u0148 bol imunitn\u00fd syst\u00e9m bakt\u00e9ri\u00ed a ich odpove\u010f na v\u00edrusov\u00fa infekciu. V d\u00f4sledku toho vznikla spolupr\u00e1ca, ktorej v\u00fdsledkom bol objav takzvan\u00fdch\u00a0\u201cgenetick\u00fdch no\u017en\u00edc\u201c \u2013 syst\u00e9mu CRISPR\/Cas9<\/strong>\u00a0a jeho vyu\u017eitie pri \u00faprave DNA. Prv\u00e1 zmienka o\u00a0tejto technol\u00f3gii bola publikovan\u00e1 v \u010dasopise Science v\u00a0roku 2012.<\/p>\n\n\n\n

Tieto genetick\u00e9 no\u017enice sa prirodzene vyskytuj\u00fa v bakt\u00e9ri\u00e1ch, v ktor\u00fdch zabezpe\u010duj\u00fa imunitn\u00fa odpove\u010f vo\u010di v\u00edrusom.<\/p>\n\n\n\n

Bakt\u00e9ria si mechanizmom CRISPR\/Cas9 zachyt\u00e1va \u00falomky DNA v\u00edrusu, ktor\u00fd ju napadol a n\u00e1sledne si vo vlastnej DNA vytvor\u00ed tzv. CRISPR \u00faseky (segmenty, obr\u00e1zok 1). V\u010faka t\u00fdmto segmentom si je bakt\u00e9ria schopn\u00e1 dan\u00fd v\u00edrus (alebo jemu podobn\u00fd v\u00edrus) zapam\u00e4ta\u0165 a v pr\u00edpade op\u00e4tovn\u00e9ho napadnutia ho vie \u013eah\u0161ie a r\u00fdchlej\u0161ie zlikvidova\u0165.<\/p>\n\n\n\n

Pre tento \u00fa\u010del si vytv\u00e1ra komplexy zlo\u017een\u00e9 z kr\u00e1tkych molek\u00fal RNA (ozna\u010den\u00fdch ako crRNA), sl\u00fa\u017eiacich na rozpoznanie v\u00edrusovej DNA a prote\u00ednov Cas, ktor\u00fdch \u00falohou je v\u00edrusov\u00fa DNA odstr\u00e1ni\u0165. RNA vznik\u00e1 v bunk\u00e1ch z DNA a je pr\u00edtomn\u00e1 pri viacer\u00fdch bunkov\u00fdch procesoch (napr\u00edklad pri tvorbe prote\u00ednov).<\/p>\n\n\n\n

\"\"
Obr\u00e1zok 1<\/strong>. Mechanizmus p\u00f4sobenia CRISPR\/Cas9 v bakteri\u00e1lnej bunke 1. Akviz\u00edcia<\/strong> \u2013 syst\u00e9m CRISPR-Cas9
uchov\u00e1va kr\u00e1tke \u010dasti DNA poch\u00e1dzaj\u00facej z v\u00edrusov, s ktor\u00fdmi sa stretol. 2. Expresia<\/strong> \u2013 pri ktorej je fin\u00e1lnym
krokom tvorba spom\u00ednan\u00fdch crRNA. 3. Interferencia<\/strong> \u2013 crRNA je vyu\u017eit\u00e1 ako vodiaca sekvencia pre prote\u00edny
Cas, ktor\u00e1 pri op\u00e4tovnej infekcii rozpozn\u00e1 v\u00edrusov\u00fa DNA a umo\u017en\u00ed t\u00fdmto prote\u00ednom dan\u00fa DNA odstr\u00e1ni\u0165.<\/figcaption><\/figure>\n\n\n\n

Vyu\u017eitie technol\u00f3gie CRISPR\/Cas9 v praxi<\/strong><\/h2>\n\n\n\n

Prv\u00e9 d\u00f4kazy o cielenom zni\u010den\u00ed DNA pomocou prote\u00ednu Cas9 rovnako, ako aj jeho schopnosti vykon\u00e1va\u0165 zmeny v gen\u00f3me (DNA) \u013eudsk\u00fdch buniek, spustili prudk\u00fd rozmach v\u00fdskumu v oblasti g\u00e9nov\u00e9ho in\u017einierstva so zameran\u00edm na CRISPR\/Cas9, ako potenci\u00e1lneho n\u00e1stroja pre \u00fapravu \u013eudskej DNA.<\/p>\n\n\n\n

Ve\u013ekou v\u00fdhodou tohto syst\u00e9mu v porovnan\u00ed s in\u00fdmi molekul\u00e1rnymi technikami s\u00fa najm\u00e4 n\u00edzke n\u00e1klady a jednoduch\u00e9 prevedenie. Princ\u00edp \u00fapravy DNA spo\u010d\u00edva v pr\u00edprave kr\u00e1tkej RNA (ozna\u010denej aj ako gRNA) tak, aby dok\u00e1zala rozpozna\u0165 nami zvolen\u00fa cie\u013eov\u00fa \u010das\u0165 (sekvenciu) v \u013eudskej DNA, ktor\u00e1 je n\u00e1sledne odstr\u00e1nen\u00e1 pomocou prote\u00ednu Cas9 (obr\u00e1zok 2).<\/p>\n\n\n\n

\"\"
Obr\u00e1zok 2<\/strong>. Rozpoznanie cie\u013eovej sekvencie DNA pomocou gRNA a prote\u00ednu Cas9.<\/figcaption><\/figure>\n\n\n\n

Po\u010diato\u010dn\u00e9 klinick\u00e9 sk\u00fa\u0161ky testovali schopnos\u0165 \u00fapravy gen\u00f3mu syst\u00e9mom CRISPR\/Cas9 na bunk\u00e1ch, ktor\u00e9 boli odobrat\u00e9 z pacienta a po \u00faprave vybran\u00fdch \u010dast\u00ed DNA boli vstreknut\u00e9 sp\u00e4\u0165 do tela.<\/p>\n\n\n\n

Prv\u00fdkr\u00e1t bola g\u00e9nov\u00e1 terapia s vyu\u017eit\u00edm CRISPR\/Cas9 testovan\u00e1 priamo v tele pacienta s genetickou mut\u00e1ciou CEP290 sp\u00f4sobuj\u00facou dedi\u010dn\u00fa poruchu zraku. Lie\u010dba je s\u00fa\u010das\u0165ou klinickej \u0161t\u00fadie, ktor\u00e1 sleduje schopnos\u0165 syst\u00e9mu CRISPR\/Cas9 efekt\u00edvne odstr\u00e1ni\u0165 mut\u00e1cie sp\u00f4sobuj\u00face Leberovu kongenit\u00e1lnu amaur\u00f3zu 10 (LCA10), na ktor\u00fa v s\u00fa\u010dasnosti neexistuje efekt\u00edvna lie\u010dba.<\/strong><\/p>\n\n\n\n

Etick\u00fd konflikt<\/h2>\n\n\n\n

V roku 2018 sa \u010d\u00ednsky lek\u00e1r He Jiankui spolu s dvoma \u010fal\u0161\u00edmi kolegami rozhodli geneticky upravi\u0165 DNA \u013eudsk\u00fdch embry\u00ed, \u010do vyvolalo pr\u00e1vny a etick\u00fd rozruch. He Jiankui p\u00f4sobiaci na Univerzite SUSTech (Southern University of Science and Technology) v \u010c\u00edne zah\u00e1jil projekt na pomoc \u013eu\u010fom, ktor\u00ed maj\u00fa probl\u00e9my s plodnos\u0165ou \u2013 konkr\u00e9tne p\u00e1rom, kde otec je HIV pozit\u00edvny a matka HIV negat\u00edvna.<\/p>\n\n\n\n

DNA embry\u00ed \u2013 konkr\u00e9tne g\u00e9n CCR5 bol upraven\u00fd vyu\u017eit\u00edm technol\u00f3gie CRISPR\/Cas9 s cie\u013eom vyvola\u0165 kompletn\u00fa rezistenciu na v\u00edrus HIV. Prv\u00e9 dve deti sa narodili v okt\u00f3bri 2018 a tretie v roku 2019. Lek\u00e1r ozn\u00e1mil, \u017ee v\u0161etky deti sa narodili zdrav\u00e9. V\u00fdskum sa v\u0161ak stretol s mas\u00edvnou kritikou a n\u00e1sledn\u00fdm suspendovan\u00edm cel\u00e9ho v\u00fdskumu. He Jiankui bol zatknut\u00fd a obvinen\u00fd z fal\u0161ovania dokumentov a neetick\u00e9ho spr\u00e1vania.<\/p>\n\n\n\n

V\u00fdzvy do bud\u00facnosti<\/h2>\n\n\n\n

Pokrok od po\u010diato\u010dn\u00e9ho objavu a charakteriz\u00e1cie CRISPR\/Cas9 syst\u00e9mu, a\u017e po jeho v\u00fdvoj v sofistikovan\u00fd, presn\u00fd a v\u0161estrann\u00fd n\u00e1stroj pre efekt\u00edvnu \u00fapravu \u013eudskej DNA, pre\u0161iel ve\u013emi r\u00fdchlym tempom. Tento n\u00e1stroj pozit\u00edvne ovplyvnil viacer\u00e9 vedeck\u00e9 i medic\u00ednske smery.<\/p>\n\n\n\n

Za menej ako osem rokov boli vyvinut\u00e9 prec\u00edzne mechanizmy (ak\u00fdm je napr\u00edklad CRISPR\/Cas9), ktor\u00e9 \u013eudstvo pribl\u00ed\u017eili k mo\u017enosti vykona\u0165 ak\u00e9ko\u013evek zmeny v DNA \u013eudsk\u00fdch buniek bez rizika ved\u013eaj\u0161\u00edch \u00fa\u010dinkov.<\/p>\n\n\n\n

Vyu\u017eitie CRISPR\/Cas9 v klinickej sf\u00e9re v\u0161ak zah\u0155\u0148a viacero prek\u00e1\u017eok s\u00favisiacich s bezpe\u010dnos\u0165ou a efekt\u00edvnos\u0165ou lie\u010debn\u00fdch postupov zalo\u017een\u00fdch na tejto technol\u00f3gii. V d\u00f4sledku toho bude pre bud\u00facnos\u0165 d\u00f4le\u017eit\u00e1 optimaliz\u00e1cia mo\u017enost\u00ed \u00fapravy \u013eudskej DNA, porozumie\u0165 d\u00f4sledkom editovania n\u00e1\u0161ho gen\u00f3mu, ako aj inovova\u0165 nov\u00e9 sp\u00f4soby dod\u00e1vania k\u013e\u00fa\u010dov\u00fdch molek\u00fal (ako napr. prote\u00edn Cas9) do buniek. D\u00f4le\u017eit\u00e9 bude aj zapojenie lek\u00e1rov, vedcov a \u010fal\u0161\u00edch zainteresovan\u00fdch str\u00e1n do tejto problematiky v d\u00f4sledku toho, aby boli tak d\u00f4le\u017eit\u00e9 objavy, ako je CRISPR\/Cas9, vyu\u017eit\u00e9 v prospech spolo\u010dnosti (napr\u00edklad na lie\u010dbu dedi\u010dn\u00fdch ochoren\u00ed).<\/p>\n\n\n\n

Zdroje<\/strong><\/p>\n\n\n\n

  1. Doudna, J. A., & Charpentier, E. (2014). The new frontier of genome engineering with CRISPR-Cas9. Science, 346(6213).<\/li>
  2. Farooq, R., Hussain, K., Tariq, M., Farooq, A., & Mustafa, M. (2020). CRISPR\/Cas9: targeted genome editing for the treatment of hereditary hearing loss. Journal of applied genetics, 61(1), 51-65.<\/li>
  3. Ledford, H. (2020). CRISPR treatment inserted directly into the body for first time. Nature, 579(7798), 185-186.<\/li>
  4. Lino, C. A., Harper, J. C., Carney, J. P., & Timlin, J. A. (2018). Delivering CRISPR: a review of the challenges and approaches. Drug delivery, 25(1), 1234-1257.<\/li>
  5. Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012). A programmable dual-RNA\u2013guided DNA endonuclease in adaptive bacterial immunity. science, 337(6096), 816-821.<\/li>
  6. Stellos, K., & Musunuru, K. (2019). Challenges and advances of CRISPR-Cas9 genome editing in therapeutics. Cardiovascular research, 115(2), e12-e14.<\/li>
  7. https:\/\/www.nobelprize.org\/prizes\/chemistry\/2020\/summary\/<\/li>
  8. https:\/\/www.science.org\/content\/article\/crispr-revolutionary-genetic-scissors-honored-chemistry-nobel<\/li>
  9. https:\/\/www.science.org\/content\/article\/untold-story-circle-trust-behind-world-s-first-gene-edited-babies<\/li>
  10. http:\/\/biochimej.univ-angers.fr\/Page2\/COURS\/Zsuite\/6BiochMetabSUITE\/4CRISPR\/1CRISPR.htm<\/li>
  11. https:\/\/ziva.avcr.cz\/ceka-nas-nastup-genove-terapie-aneb-homo-sapiens-gmo.html<\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

    V\u00f4bec po prv\u00fdkr\u00e1t v hist\u00f3rii boli tento rok ocenen\u00e9 Nobelovou cenou za ch\u00e9miu dve \u017eeny. Americk\u00e1 vedky\u0148a Jennifer Doudna, profesorka na Kalifornskej univerzite v meste Berkeley a franc\u00fazska vedky\u0148a Emmanuelle Charpentier, profesorka, geneti\u010dka, mikrobiologi\u010dka a riadite\u013eka \u00fastavu Maxa Plancka pre infek\u010dn\u00fa biol\u00f3giu. Spolo\u010dne objavili met\u00f3du na cielen\u00fa \u00fapravu genetickej inform\u00e1cie (DNA) v rastlinn\u00fdch, \u017eivo\u010d\u00ed\u0161nych, ako aj v \u013eudsk\u00fdch bunk\u00e1ch.\u00a0Hlavn\u00fdm […]<\/p>\n","protected":false},"author":1,"featured_media":2414,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"content-type":"","inline_featured_image":false,"footnotes":""},"categories":[1],"tags":[63],"place-taxonomy":[],"class_list":["post-2413","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-uncategorized-sk","tag-zaujimavosti"],"acf":[],"views":2708,"_links":{"self":[{"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/posts\/2413","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/comments?post=2413"}],"version-history":[{"count":4,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/posts\/2413\/revisions"}],"predecessor-version":[{"id":1352810,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/posts\/2413\/revisions\/1352810"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/media\/2414"}],"wp:attachment":[{"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/media?parent=2413"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/categories?post=2413"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/tags?post=2413"},{"taxonomy":"place-taxonomy","embeddable":true,"href":"https:\/\/dnaera.com\/ro\/wp-json\/wp\/v2\/place-taxonomy?post=2413"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}