CRISPR/Cas9, a bacterial immune system became the most widely used tool for genome engineering

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68378050%3A_____%2F19%3A00503188" target="_blank" >RIV/68378050:_____/19:00503188 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

CRISPR/Cas9, a bacterial immune system became the most widely used tool for genome engineering

Popis výsledku v původním jazyce

The CRISPR/Cas system has revolutionized the way we perform genomic manipulations in a wide range of cell and model organisms today and has great potential for therapeutic application. Due to its simple design and affordability it has become a versatile research tool, used in many diverse applications ranging from genomic manipulations to study the function of individual genes to high-throughput screens covering the whole genome. The major CRISPR/Cas system currently used is the RNA-guided Cas9 endonuclease which originates from Streptococcus pyogenes (SpCas9). The original CRISPR/Cas9 is composed of three components: two short guide RNAs (a crRNA which defines the target site on DNA and a tracrRNA connecting the crRNA) and the Cas9 protein. Following complex formation of the guide RNAs with the Cas9 protein, the endonuclease activity becomes activated, allowing the introduction of site-specific DNA double strand breaks for genomic manipulations. The specificity of CRISPR/Cas9 is given by a 20 nt sequence of the guide RNA, which has to be complementary to the DNA target site, and the requirement of a PAM sequence recognized by the Cas9 protein. The PAM sequence within the target DNA for Cas9 is NGG. Although the original CRISPR/Cas9 system has proven to be a very powerful and robust tool for genome engineering, it does have some drawbacks with the main concerns being the specificity, e.g. undesired side effects on off-target sites with similar sequences to the on-target site, and the restrictions due to the requirement of the PAM sequence, which limits the number of accessible target loci within the genome. In this mini review we aim to give an overview of the SpCas9 system and the improved versions of the original Cas9 protein addressing the aforementioned complications.

Název v anglickém jazyce

CRISPR/Cas9, a bacterial immune system became the most widely used tool for genome engineering

Popis výsledku anglicky

The CRISPR/Cas system has revolutionized the way we perform genomic manipulations in a wide range of cell and model organisms today and has great potential for therapeutic application. Due to its simple design and affordability it has become a versatile research tool, used in many diverse applications ranging from genomic manipulations to study the function of individual genes to high-throughput screens covering the whole genome. The major CRISPR/Cas system currently used is the RNA-guided Cas9 endonuclease which originates from Streptococcus pyogenes (SpCas9). The original CRISPR/Cas9 is composed of three components: two short guide RNAs (a crRNA which defines the target site on DNA and a tracrRNA connecting the crRNA) and the Cas9 protein. Following complex formation of the guide RNAs with the Cas9 protein, the endonuclease activity becomes activated, allowing the introduction of site-specific DNA double strand breaks for genomic manipulations. The specificity of CRISPR/Cas9 is given by a 20 nt sequence of the guide RNA, which has to be complementary to the DNA target site, and the requirement of a PAM sequence recognized by the Cas9 protein. The PAM sequence within the target DNA for Cas9 is NGG. Although the original CRISPR/Cas9 system has proven to be a very powerful and robust tool for genome engineering, it does have some drawbacks with the main concerns being the specificity, e.g. undesired side effects on off-target sites with similar sequences to the on-target site, and the restrictions due to the requirement of the PAM sequence, which limits the number of accessible target loci within the genome. In this mini review we aim to give an overview of the SpCas9 system and the improved versions of the original Cas9 protein addressing the aforementioned complications.

Druh

C - Kapitola v odborné knize

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/LO1220" target="_blank" >LO1220: CZ-OPENSCREEN: Národní infrastruktura chemické biologie</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název knihy nebo sborníku

Advances in Chemical Biology

ISBN

978-80-88011-03-3

Počet stran výsledku

9

Strana od-do

92-103

Počet stran knihy

210

Název nakladatele

OPTIO CZ

Místo vydání

Praha

Kód UT WoS kapitoly

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