CRISPR-mediated activation in E. coli [20]. There are

CRISPR-mediated gene activation, called
CRISPRa, uses dCas9 fusion proteins to recruit transcription activators. A
fusion of dCas9 with the ?-subunit of the E. coli Pol allowed assembly of the
holoenzyme at a target promoter for gene activation in E. coli 20. There are currently limited
reports on CRISPRa in bacteria, and more work is needed to achieve robust and
consistent gene activation in bacteria 5.

CRISPR–dCas9 can target several genes
simultaneously by using multiple sgRNAs. Recently, a method for simultaneous
repression and activation of genes was established using scaffold RNAs (scRNAs)
21.ACUTE MYELOID
LEUKEMIA (AML)22 identified additional therapeutic targets in
acute myeloid leukemia (AML), they optimized a genome-wide clustered regularly
interspaced short palindromic repeats (CRISPR) screening platform and use it to
identify genetic vulnerabilities in AML cells. They identified 492 AML-specific
cell-essential genes, including several established therapeutic targets such as
DOT1L, BCL2, and MEN1, and many other genes including clinically actionable
candidates, validated selected genes using genetic and pharmacological inhibition,
and chose KAT2A as a candidate for downstream study. KAT2A inhibition
demonstrated anti-AML activity by inducing myeloid differentiation and
apoptosis, and suppressed the growth of primary human AMLs of diverse genotypes
while sparing normal hemopoietic stem-progenitor cells, results proposed that
KAT2A inhibition should be investigated as a therapeutic strategy in AML and
provide a large number of genetic vulnerabilities of this leukemia that can be
pursued in downstream studies.23  Using the IDH2 R140Q mutation as a model,
presented a new effective methodology here using the RNA guided clustered
regularly interspaced short palindromic repeats (CRISPR)-Cas9 system to
reproduce or remove AML associated mutations in or from human leukemic cells,
respectively, via introduction of a DNA template at the endogenous gene locus
via homologous recombination. Our technology represents a precise way for AML
modeling to gain insights into AML development and progression and provides a
basisfor
future therapeutic approaches. Isocitrate
dehydrogenases (IDHs) are digestive enzymes that catalyse the oxidative
decarboxylation of isocitrate, producing alpha-ketoglutarate (a-ketoglutarate)
and CO2. Mutations in IDH1/2 genes occur frequently in AML patients, and IDH2
R140Q has been shown to be the most frequent IDH mutation in AML 24. IDH2 R140Q has
been identified as a key driver mutation in a transgenic mouse model,
supporting its relevance as a therapeutic target for the treatment of human AML
25.Despite intense
research into acute myeloid leukemia (AML) during past decades, the majority of
AML patients still die from their disease. Thus, there is a high need for new
AML therapies. AML is a heterogeneous disease harboring a multitude of genetic
and epigeneticchanges,
and it is highly likely that the various AML subtypes require different
targeted therapeutic approaches 23.

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