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Keywords
Abbreviations
Introduction
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Figure 1: Mechanisms of CRISPR/Cas9 mediated bacterial immune-defense. Phase 1: Immunization process: the CRISPR locus contains a
series of short repeated sequences that are separated by non-repetitive spacer sequences derived from foreign genetic elements. Invading foreign
DNA from viruses and phages is processed by Cas nuclease into small DNA fragments, which are then incorporated into the CRISPR locus of host
genomes as new spacers. Phase 2: Immunity process: In response to viruses and phage infections, the CRISPR repeat-spacer array is transcribed
into a pre-crRNA and processed into mature crRNAs, which guide Cas to cleave foreign DNA.
Citation: Zhao Y, Ying Y, Wang Y (2014) Developing CRISPR/Cas9 Technologies for Research and Medicine. MOJ Cell Sci Report 1(1): 00006.
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Repurposing CRISPR/Cas9
Genetic/Epigenetic Modulation
The CRISPR/Cas9 system has the potential to regulate
endogenous gene expression. Catalytically inactive or dead Cas9
(dCas9) bearing mutations at the two cleavage sites renders Cas9
unable to cut DNA, but it can still be recruited by gRNAs to target
specific DNA sites within the genome [14,21,67]. Therefore, dCas9
with heterologous effector domain functions can be recruited
to specific genome loci, thus modulating the genetic/epigenetic
states (Figure 2B). dCas9 fusions to a transcriptional activation
domain (e.g. VP64 and the p65 subunit of nuclear factor kappa B;
NF-B) or a transcriptional repression domain (e.g. the Krppelassociated box (KRAB) domain) implements precise and stable
transcriptional control of target genes [68-72]. For example,
single or multiple gRNAs can direct a VP64 transcriptional
activation domain fused with a dCas9 protein to activate the
expression of several endogenous genes [68,69]. An alternative
CRISPR/Cas9
TALE
ZF
Mega nuclease
RNA-DNA
Protein-DNA
Protein-DNA
Protein-DNA
1. Genome editing;
2. Genetic and epigenetic modulation;
1. Genome editing;
2. Genetic modulation;
Citation: Zhao Y, Ying Y, Wang Y (2014) Developing CRISPR/Cas9 Technologies for Research and Medicine. MOJ Cell Sci Report 1(1): 00006.
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Citation: Zhao Y, Ying Y, Wang Y (2014) Developing CRISPR/Cas9 Technologies for Research and Medicine. MOJ Cell Sci Report 1(1): 00006.
Acknowledgement
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Citation: Zhao Y, Ying Y, Wang Y (2014) Developing CRISPR/Cas9 Technologies for Research and Medicine. MOJ Cell Sci Report 1(1): 00006.