According to the characteristics of different cell lines, appropriate transfection methods (virus
liposome transfection, and electroporation) will be selected to transfer gRNA sequences and Cas9
cells, and antibiotic screening with different duration will be carried out according to different
methods used. After antibiotic screening, single-cell clones will be generated. Positive clones that
successfully knocked out will be validated by target site amplification and sequencing. Final
the homozygous KO cell clones, related data and project reports.
KO cell bank
3000 KO cell lines Starting from $1780 Deliver in 1 week!
Ubigene's KO cell bank covers thousands of genes, including 8 signal pathways, 10+ drug
development sectors, 100+ common diseases and popular research fields (such as m6A, ferroptosis
and exosomes). Search your target gene below to find out if there is a KO cell line in stock.
Knockout Cell Service
Various types of cells including tumor cell lines, regular cell lines,
Human Breast Cancer Cell Line(MCF7)Mouse insulinoma β cell line(NIT-1)Human Breast Cancer Cell Line(JIMT-1)Human breast cancer cell line(T-47D)Human pancreatic cancer cell line(BxPC-3)Mouse Acinar Pancreatic Cell Line(266-6)Human Prostate Cancer Cell Line(VCaP)Human Pancreatic Carcinoma Cell Line(MIA PaCa-2)Mouse medullary breast cancer cell line(E0771)Mouse pancreatic cancer cell line(Pan02)Human Metastatic Pancreatic Adenocarcinoma Cell Line(AsPC-1)Human Breast Adenocarcinoma Cell Line(SK-BR-3)Human Pancreatic Carcinoma Cell Line(PANC-1)Rat Breast Cancer Cell Line(4T1)Human Breast Cancer Cell Line(ZR-75-1)Human Breast Cancer Cell Line(MDA-MB-231)
Guide RNAs target introns at both sides of exon 2 and the number of bases in exon 2 is not a
which can cause frame-shift mutation.
Guide RNA targets the exon, and the base number of deletion is not a multiple of 3. After
frame-shift mutation would cause gene knockout.
Complete removal of the coding sequence to achieve gene knockout.
Work Flow and Validation
Chinese hamster ovary (CHO) cells have been used as host cells in the
range of recombinant therapeutic proteins,
antibodies and Fc-fusion proteins. Host cell proteins (HCP) represent impurities that must be
therapeutic formulations because of their potential risks for immunogenicity. While the majority
are effectively removed in typical downstream purification processes, clearance of a small
population of HCP
remains challenging. Using the CRISPR/Cas9 system, Anxa2-, and Ctsd-knockout CHO cell lines were
established, and this study confirmed the complete elimination of the corresponding HCP in cell
Importantly, all knockout cell lines showed similar growth and viability to those of the
8 days of cultivation. Thus, knockout of unrequired genes can reduce contamination with HCP in
production of recombinant therapeutic
（a）The constructed sgRNA targeted a unique sequence in exon 4 of the Anxa2
amplicon sequence was analyzed in both directions.
（b）The constructed sgRNA targeted the unique sequence in exon 2 of the Ctsd
amplicon sequence was analyzed in both directions.
Characterization of protein expression from CHO knockout cell lines by
blotting analysis. (a) Anxa2-knockout cell lines. (b) Ctsd-knockout cell lines. Cell
supernatants and cell lysates were subjected to SDS-PAGE. Total proteins were detected by CBB
blotting analysis of each protein was performed using respective capture and detection
asterisk indicates the nonspecific band. The double asterisk indicates what is likely a fragment
Annexin A2 and cathepsin D were not detected in the cell culture supernatants
the corresponding knockout cell lines, suggesting successful exclusion of these impurities from
therapeutic protein production. In addition, no truncated HCP was observed in the protein
the Anxa2 and Ctsd knockout cell lines.
Fukuda, N., Senga, Y., & Honda, S. (2019). Anxa2‐and Ctsd‐knockout CHO
diminish the risk of contamination with host cell proteins. Biotechnology progress, e2820.
Selected KO-cited papers
STING1 Knockout in HeLa cell line——prevention and treatment of viral diseases
STING1 Knockout in HeLa cell line——prevention and treatment of viral diseases IF=32.4 Immunity
to study the regulation mechanism of cGAS activity in order to prevent and treat
viral diseases, a research team leading by Prof. Zhao in Shandong University used the HeLa cell line with STING1 gene knockout constructed by Ubigene as the key cell model, and found out that the two conformations of DNA (B-DNA and
Z-DNA) had different affinity with cGAS; The endogenous metabolism of small
molecules spermine, spermidine and polyamine catabolism key enzyme SAT1 regulates
cGAS activity by inducing DNA conformational transition. This research reveals a
additional mechanism for preventing abnormal cytoDNA recognition and providing
promising therapeutic targets for the treatment of diseases involving improper cGAS
Polyamine metabolism regulates cGAS activity pattern by controlling Z-DNA
TP53 Knockout in HCT116 cell line——Apoptosis
TP53 Knockout in HCT116 cell line——Apoptosis IF=16.6 Nature Communications
Published by: Xiangya Hospital, Central South University
In this study, the HCT116 cell line with TP53 gene knocked out constructed by Ubigene was
By analyzing the crystal structure of the complex of p53 and the anti-apoptotic
protein BCL-2, and combining biochemical and cellular experiments, this study
revealed a new mechanism of p53 interacting with BCL-2 protein and promoting
apoptosis, that is, p53 forms a complex with BCL-2 by directly occupying the
BH3-binding pocket of BCL-2, and antagonizes BCL-2 activity by releasing
pro-apoptotic BCL-2 family proteins located in the pocket, thereby promoting
apoptosis.These structural and functional data
provide a new idea for further understanding the complex regulatory mechanism of
p53-mediated mitochondrial apoptosis, and also provide an important basis for
developing anticancer therapeutic strategies that target protein-protein
interactions to activate apoptosis. View
Mutations in p53 interacting with BCL-2 reduce apoptosis
ZNF432 knockout U2OS cell line——Ovarian cancer drug resistance
Nucleic Acids Research
ZNF432 knockout U2OS cell line——Ovarian cancer drug resistance IF=14.9 Nucleic Acids Research
ZNF protein was confirmed as a key factor regulating the genome integrity of
mammalian cells. In order to explore the possibility that ZFP can be used as an
effector of DNA repair based on homologous recombination (HR), Jean Yves Masson team of Laval University used the ZNF432 knockout U2OS cell line constructed by Ubigene to carry out a series of experiments, and found that ZNF432 deletion in cancer cells
would accelerate DNA repair, lead to the weakening of PARPi effect, and make ovarian
cancer cells develop drug resistance, confirming that ZNF432 is a new HR inhibitor,
which successfully broadened the new way to study the efficacy of PARPi. View details>>
Regulation of ZNF432 expression affects drug resistance of ovarian cancer cells
SNORD17 Knockout in HepG2 cell line——liver tumors
Cell death & differentiation
SNORD17 Knockout in HepG2 cell line——liver tumors IF=12.4 Cell death & differentiation
Published by: Hepatic Surgery Center, Tongji Hospital
This study reveals the regulatory role of small nucleolar RNA SNORD17 and p53
pathway in hepatocellular carcinoma, which provides a new potential target for the
treatment of hepatocellular carcinoma . The researchers utilized SNORD17 knockout Hep G2 cell line (constructed by Ubigene) as
the key cell model. After applied with in vitro and in vivo tests, they found out
that in HCC cell lines, the knockout of SNORD17 gene can significantly inhibit cell
proliferation, clone formation and G1/S phase transition. View
p53 mediates SNORD17 to affect HCC cell growth
Pik3r1 knockout RAW264.7 cell line - Osteoporosis
Pik3r1 knockout RAW264.7 cell line - Osteoporosis IF=11.4 Redox Biology
Published by: Orthopedics Research Institute, Zhejiang University
This article explores a new mechanism for GSTP1 affecting the osteoclast
cell-related bone homeostasis through combining with a large number of in vivo and
in vitro experiments, based on the Pik3r1 knockout RAW264.7 cell model constructed by Ubigene.
It was the first time to interpret that the cell fate of osteoclasts is determined
by S-glutathionylation via a redox-autophagy which is mediated by GSTP1. View
SiNPs induces CASA mechanism
KO cells are often used as key models for studying gene function, disease mechanisms, screening drug targets,
. Based on the rich gene editing experience, Ubigene has established a stock of over 3500 KO cells, which can meet most scientific research needs. If you would
like to get a KO cell line in-stock/customized, please contact
References: Zhao C, Ma Y, Zhang M, et al. Polyamine metabolism controls B-to-Z DNA transition to orchestrate
DNA sensor cGAS activity[J]. Immunity, 2023, 56(11): 2508-2522. e6. Wei H, Wang H, Wang G, Qu L, Jiang L, Dai S, Chen X, Zhang Y, Chen Z, Li Y, Guo M, Chen Y.
Structures of p53/BCL-2 complex suggest a mechanism for p53 to antagonize BCL-2 activity. Nat
Commun. 2023 Jul 18;14(1):4300. O’Sullivan J, Kothari C, Caron M C, et al. ZNF432 stimulates PARylation and inhibits DNA
resection to balance PARPi sensitivity and resistance[J]. Nucleic Acids Research, 2023, 51(20):
11056-11079. Zhu Y, Zhang Y, Fan Z, et al. Silica Nanoparticles Trigger Chaperone HSPB8‐Assisted Selective
Autophagy via TFEB Activation in Hepatocytes[J]. Small, 2023, 19(5): 2204310. Liang J, Li G, Liao J, et al. Non-coding small nucleolar RNA SNORD17 promotes the progression of
hepatocellular carcinoma through a positive feedback loop upon p53 inactivation[J]. Cell Death &
Differentiation, 2022, 29(5): 988-1003.
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