CRISPR Gene Editing in Stem Cells

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Table 1. Examples of CRISPR/Cas9 Being Used for Cell Biology Research
Organelle CRISPR/Cas9 Target Finding Refs
Microtubule CRISPR/Cas9 generation of mutant
fl ies by deleting a linker region in the
centrosome protein CP190
Identifi ed a centrosome and
microtubule-targeting region in
CP190 for spindle localization;
deletion of linker region altered
spindle morphology and led to DNA
segregation errors
[114]
Mitochondrion CRISPR/Cas9 knockout of copper
transporting ATPase ATP7A in
mouse 3T3-L1 cells and in
fi broblasts from patients with
Menkes Disease (MD)
ATP7A dysfunction damages
mitochondrial redox balance
[115]
CRISPR/Cas9 knockout of
FASTKD2, a RNA-binding protein of
the FAS-activated serine/threonine
kinase family
Defective processing and
expression of mitochondrial RNA;
cellular respiration damage with
depressed activities of respiratory
complexes
[116]
CRISPR/Cas9-mediated repair of
ARID5B motif of rs1421085 in
primary adipocytes from a patient
carrying the risk allele
IRX3 and IRX5 repression restored;
browning expression programs
activated; thermogenesis restored
[117]
CRISPR/Cas9-based genetic
screen to study cell proliferation
suppression due to inhibition of
mitochondrial electron transport
chain (ETC)
Identifi ed cytosolic aspartate
aminotransferase (GOT1) as key
gene; GOT1 loss-of-function kills
cells upon ETC inhibition
[118]
Endoplasmic
reticulum
CRISPR/Cas9 knockout of ATF4 or
NLRP1
NLRP1 upregulated during severe
ER stress; ATF4 binds and activates
NLRP1 promoter during ER stress
[119]
CRISPR/Cas9-mediated deletion of
transmembrane endoribonuclease
Ire1/ in HEK293 cells
Ire1/ forms a complex with the
Sec61 translocon to cleave its
mRNA substrates; disruption of
Ire1/ complex reduced cleavage of
ER-targeted mRNA
[120]
Centrosome CRISPR/Cas9 dual-sgRNA to
generate a null abnormal spindle
(asp) allele by excising a 750-bp
fragment that included the
promoter, 50 UTR, and the first exon
in Drosophila neuroblasts
Asp null mutations cause spindle
defects in neuroblasts; Asp
regulated by Drosophila
melanogaster calmodulin (CaM) to
crosslink spindle microtubules
[121]
Lysosome Generation of Niemann-Pick type C
1 (NPC1)-deficient cell line using
CRISPR/Cas9
NPC1 moves cholesterol across
lysosomal glycocalyx
[122]
Ribosome CRISPR/Cas9 knockout of
nonessential gene of ribosomal
protein eS25 (RPS25) in Hap1 cell
line; RPS25-SNAP (mutant O6-
alkylguanine DNA alkyl-transferase)
transgene was transduced into
RPS25-KO Hap1 cells to be the only
source of the protein
Demonstrated an approach to
create fluorescently labeled 40S
ribosomal subunits from human
cells; studied kinetics of the 40S
subunit recruitment to the hepatitis
C virus (HCV) internal ribosome
entry site (IRES)
[123]
Golgi apparatus Genome-wide CRISPR/Cas9 loss-
of-function screen to identify host
targets required for Staphylococcus
aureus toxin alpha hemolysin (/HL)
susceptibility in human myeloid cells
Identifi ed new proteins (SYS1,
ARFRP1, and TSPAN14) in
regulating presentation of ADAM10
on the plasma membrane post-
translationally; cells lacking
sphingomyelin synthase 1 (SGMS1)
resist / HL intoxication
[124]
Trends in Cell Biology, November 2016, Vol. 26, No. 11 879