THP-1-CAS9(THP-1-CAS9)
Catalog#: YC-D011-Cas9-H
Size: 1*10^6
Instruction:
A human leukemic cell line (THP-1) cultured from the blood of a boy with acute monocytic leukemia. Since its establishment in 1980, THP-1 cells have been widely used in the research of monocyte and macrophage related mechanisms, signaling pathways, nutrient and drug transportation. The morphological and functional characteristics of THP-1 are very similar to human primary monocytes (including cell differentiation markers). Compared with human peripheral blood monocytes (PBMC), THP-1 is easier to be cultured and has a more consistent background. Therefore, THP-1 is a commonly used acute monocytic leukemia cell line in various laboratories, and it is an ideal tool for studying immunity and inflammation.
Application of THP-1: macrophage and inflammation model
THP-1 can be differentiated into M1/M2 macrophages and release
corresponding cytokines.
· M1 macrophage
polarization：
THP-1 can be induced to differentiate into macrophages by Phorbol 12-myristate 13-acetate
(PMA), and then M1 polarization can be induced by lipopolysaccharide (LPS) and IFN -γ,
releasing TNF -α, IL-6 and other cytokines. This is a typical inflammatory model.
· M2 macrophage
polarization：
M2 polarization can be induced by IL-4, IL-13 and
macrophage colony-stimulating factor (M-CSF). TGF - β, IL-10 and other inhibitory cytokines
can be released. This is similar to the process of tissue repair and reconstruction in the
late stage of inflammation.
· Atherosclerotic
inflammation model：
Under the action of oxidized low-density lipoprotein
(ox-LDL), macrophages can further become foam cells. This is a pathological cell in
atherosclerotic plaques and is a chronic inflammation model.
CRISPR-U™ for efficient gene editing in THP-1 cells
THP-1 is a near-tetraploid suspension cell, and the success rate of THP-1 is very low by
conventional gene-editing methods. CRISPR/Cas9 is widely used to construct gene-editing
THP-1 model because of its simple, high efficiency and low toxicity. CRISPR-U™, developed by
Ubigene, is more efficient than general CRISPR/Cas9 in double-strand breaking, and CRISPR-U™
can greatly improve the efficiency of homologous recombination, easily achieve knockout
(KO), point mutation (PM) and knockin (KI). Ubigene can customize the gene-editing THP-1 cell line and other monocytes that you are
interested in, as well as generate various genes overexpression in THP-1 cell line.
THP-1 Cell Line Gene-editing Services：
Technical advantages
Exclusive innovation, 10 times more efficient in gene-editing.
Successfully edit genes on more than 100 types of cell lines.
Easily generate knockout (KO), point mutation (PM) and knockin (KI) in vitro and in
vivo.
CRISPR-U™ offers a 100% mutation guarantee. No mutation, no charge!Gene Knockout
Point mutation (PM)
CRISPR-U™ Point Mutation THP-1 Cell Line： THP-1 cell line would be efficiently co-transfected with gRNA, Cas9 and ssODN.
After drug screening, single clones would be generated. Positive clones would be
validated by sequencing.
· Disease model
generation
ssODN carrying point mutaion which replaces the WT sequence
by HDR.
· Disease model rescuing
ssODN carrying WT sequence which replaces the mutated site
by HDR.
Case Study:
CRISPR/Cas9 mediated Chronic granulomatous disease
(CGD) THP-1 cell line model is helpful to develop better disease treatments
Chronic granulomatous disease (CGD) is a rare X-linked genetic disease. Due to
the mutation or deficient of CYBB gene, macrophages lack nicotinamide adenine
dinucleotide phosphate (NADPH) oxidase, and cannot produce hydrogen peroxide to
effectively kill the invading microorganisms. This usually leads to serious
repeated infections caused by bacteria, fungi and other microorganisms. Some
researchers used CRISPR/Cas9 technology to knockout CYBB gene and generate point
mutation c.90c>G (found in a CGD patient) in THP-1 cells, and successfully
constructed the CGD model. Compared with wild-type THP-1 cells, two KO clones
(#3 and #27) and a point mutation clone (#2, c.90c > G) showed decrease in H2O2
level after PMA and LPS induction, and a significant increase in IL-1β, TNF-α
and IL-6 release, which was consistent with the behavior of macrophages in
CGD.This CGD model provides a powerful tool for disease study and will help
to develop better treatments.
Compared with wild-type THP-1 cells, two KO clones (#3 and
#27) and a point mutation clone (#2, c.90c > G) showed decrease in H2O2 level
after PMA and LPS induction, and a significant increase in IL-1β, TNF-α and IL-6
release, which was consistent with the behavior of macrophages in CGD. This CGD
model provides a powerful tool for disease study and will help to develop better
treatments.
Reference：
Benyoucef A, Marchitto L, Touzot F. CRISPR gene-engineered CYBBko THP-1 cell
lines highlight the crucial role of NADPH-induced reactive oxygen species for
regulating inflammasome activation[J]. Journal of Allergy and Clinical
Immunology, 2020.
Gene knock in
