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

Result code in 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>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

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

Original language description

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.

Czech name

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Czech description

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Type

C - Chapter in a specialist book

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

<a href="/en/project/LO1220" target="_blank" >LO1220: CZ-OPENSCREEN: National infrastructure for chemical biology</a><br>

Continuities

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

Publication year

2019

Confidentiality

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

Book/collection name

Advances in Chemical Biology

ISBN

978-80-88011-03-3

Number of pages of the result

9

Pages from-to

92-103

Number of pages of the book

210

Publisher name

OPTIO CZ

Place of publication

Praha

UT code for WoS chapter

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