Knockout Cell Lines

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Knockout Cell Line

According to the characteristics of different cell lines, appropriate transfection methods (virus transduction, liposome transfection, and electroporation) will be selected to transfer gRNA sequences and Cas9 protein into cells, and antibiotic screening with different duration will be carried out according to different transfection methods used. After antibiotic screening, single-cell clones will be generated. Positive clones that are successfully knocked out will be validated by target site amplification and sequencing. Final deliverables will be the homozygous KO cell clones, related data and project reports.
KO cell bank
 3000 KO cell lines    Starting from $1780    Deliver in 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
Cell type Various types of cells including tumor cell lines, regular cell lines, IPS/ES cell lines
Service type Single / Multiple Genes Knockout
Deliverables Cell pool/Single-cell clone
Turnaround/Price Speedy turnaround as fast as 4 weeks!     Contact Us

200+Successful
Gene-editing Cell Line Types

Respiratory System
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Chinese hamster lung cells(V79)Human hypopharyngeal carcinoma cell line(FaDu)Human Bronchial Epithelial Cell Line(16HBE)Human Bronchial Epithelial Cell Line(BEAS-2B)Human Non-small Cell Lung Carcinoma Cell Line(HCC827)Human Non-small Cell Lung Carcinoma Cell Line(NCI-H1299)Human Lung Squamous Cell Carcinoma Cell Line(NCI-H226)Human lung squamous cell carcinoma cell line(SK-MES-1)Human Lung Cancer Cell Line(NCI-H520)Human Lung Cancer Cell Line(Calu-1)Human Lung Cancer Cell Line(A549)
Circulatory System
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Mouse Myoblast Cell Line(C2C12)Human Coronary Artery Endothelial Cell line(HCAEC)Rat Cardiac Myocytes(HL-1)Rat Cardiac Myocytes(H9C2)
Endocrine System
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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)
Brain and Nervous System
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Human glioma cell line(U251MG)Mouse microglia cell line(BV2)Immortalize Human Microvascular Endothelial Cell Line(hCMEC/D3)Mouse Anterior Parietal Bone Cell Line(MC3T3-E1 Subclone 14)Human glioblastoma cell line(U-87 MG)Rat Glioblastoma Cell Line(C6)Mouse neuroblastoma cell line(Neuro-2a)Human Neuroblastoma Cell Line(SK-N-SH)
Blood and lymphatic System
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Human B lymphoma cell line(Su-DHL-4)Human B Cell Lymphoma Cancer Cell Line OCI-LY3(OCI-LY3)Human B Cell Lymphoma Cancer Cell Line(U2932)Human Acute Non-B Non-T Lymphocytic Leukemia Cell Line(Reh)Human myelogenous leukemia cell line(K-562)Human T lymphocyte cell line(Jurkat, Clone E6-1)Rat Basophil Leukemia Cell Line(RBL-2H3)Human Monocytic Cell Line(THP-1)Porcine alveolar macrophage cell line(3D4/21)Mouse Macrophage Cell Line(RAW264.7)
Urinary System
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Mouse prostate cancer cell line(RM-1)Rat adrenal pheochromocytoma cell line(PC-12)Distal nephron cell line(Distal nephron cell line(JU4s))Dog Kidney Cell Line(MDCK)Human prostate cancer cell line(22RV1)Human Embryonic Kidney Cell Line(293T)Human Embryonic Kidney Cell Line(HEK293)Human Bladder Transitional Cell Carcinoma Cell Line(T24)Human bladder carcinoma cell line(5637)Human bladder carcinoma cell line(TCCSUP)
Digestive System
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Human colon adenocarcinoma cell line(LS174T)Porcine intestinal epithelial cell line(IPEC-J2)Human hepatoma cell line(HepaRG)Mouse Hepatocarcinoma Cell Line(Hepa 1-6)Mouse Hepatocarcinoma Cell Line(H22)Human Hepatoma Cell Line(SMMC-7721)Human renal carcinoma cell line(ACHN)African green monkey kidney cell(Vero)Human renal cell carcinoma cell line(786-0)Human Esophageal Squamous Carcinoma Cell Line(KYSE-30)Human Esophageal Squamous Carcinoma Cell Line(KYSE-150)Human gastric cancer cell line(AGS)Human Gastric Cancer Cell Line(SGC-7901)Human Gastric Cancer Cell Line(HGC-27)Human hepatobiliary cancer cell line(RBE)Human hepatocellular carcinoma cell line(HuH-7)Human Hepatoma Cell Line(Hep3B)Human liver cancer cell line(Hep G2)Human Normal Hepatocytes Cell line(L-02)Human colon carcinoma cell line(T84)Human colorectal adenocarcinoma cell line(Caco-2)Human colorectal adenocarcinoma cell line(NCI-H716)Human colon adenocarcinoma cell line(DLD-1)Murine colorectal carcinoma cell line(CT26.WT)Murine Colorectal Carcinoma Cell Line(MC38)Human caucasian colon adenocarcinoma cell line(COLO 205)Human colon carcinoma cell line(RKO)Human Colon Cancer Cell Line(HT-29)Human Colon Cancer Cell Line(SW620)Human Colon Cancer Cell Line(SW480)Human Colon Cancer Cell Line(HCT 116)
Skeleton, Articulus, Soft Tissue, Derma System
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Mouse osteoid cell line(MLO-Y4)Ameloblastoma(hTERT-AM)Mouse squamous carcinoma cell line(SCC7)Mouse myeloma cell line(Sp2/0-Ag14)Human skin squamous carcinoma cell line(A431)Murine melanoma cell line(B16-F10)Human Melanoma Cell Line(M14)Human malignant melanoma cell line(A-375)Human fibrosarcoma cell line(HT-1080)Human bone osteosarcoma epithelial cell line(U-2 OS)Human Osteosarcoma Cell Line(MG-63)
Ocular, Otolaryngologic and Oral System
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Human retinal pigment epithelial cell line(ARPE-19)Human choroidal melanoma cell line(OCM-1)Human oral squamous carcinoma cell line(HSC3)Human Nasopharyngeal Carcinoma Cell Line(C666-1)Human Nasopharyngeal Carcinoma Cell Line(CNE2Z)Human nasopharyngeal carcinoma cell line(NPC-43)Rat Muller Cell Line(rmc-1)
Reproductive System
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Human ovarian cancer cell line(OVCAR-3)Human Villous Trophoblast(HTR-8/SVneo)Human Ovarian Adenocarcinoma Cell Line(CAOV3)Mouse Testicular Stromal Cell Line(TM3)Mouse Pituitary Cell Line(Lbetat2)Human Ovarian Cancer Cell Line(SK-OV-3)Chinese Hamster Ovary Cell Line(CHO-K1)Mouse Embryonic Fibroblasts(NIH/3T3)Human Cervical Carcinoma Cell Line(HeLa 229)Human Cervical Carcinoma Cell Line(HeLa)
Stem Cell
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Stem Cell(H9)Human Embryonic Stem Cell(H1)Induced Pluripotent Stem Cell(ipsc)
CRISPR-U™ Technical advantage
Higher cutting and homologous recombination efficiency, 10-fold increase in gene-editing efficiency

Higher cutting and homologous recombination efficiency, 10-fold increase in gene-editing efficiency

A large number of success case, successfully achieving over 5000 gene edits in over 200 cell line types

A large number of success case, successfully achieving over 5000 gene edits in over 200 cell line types

Wide application, easily achieving gene knockout / point mutation / knock-in

Wide application, easily achieving gene knockout / point mutation / knock-in

Knockout Strategies
Short fragment removalShort fragment removal

Guide RNAs target introns at both sides of exon 2 and the number of bases in exon 2 is not a multiple of 3, which can cause frame-shift mutation.

Frame-shift mutationFrame-shift mutation

Guide RNA targets the exon, and the base number of deletion is not a multiple of 3. After knockout, frame-shift mutation would cause gene knockout.

Large fragment removalLarge fragment removal

Complete removal of the coding sequence to achieve gene knockout.

CRISPR-U KO Strategy diagram
Work Flow and Validation
Knockout Cell Line Work Flow and ValidatioKnockout Cell Line Work Flow and Validatio
Case Study
Chinese hamster ovary (CHO) cells have been used as host cells in the production of a range of recombinant therapeutic proteins, including monoclonal antibodies and Fc-fusion proteins. Host cell proteins (HCP) represent impurities that must be removed from therapeutic formulations because of their potential risks for immunogenicity. While the majority of HCP impurities 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 successfully established, and this study confirmed the complete elimination of the corresponding HCP in cell lysates. Importantly, all knockout cell lines showed similar growth and viability to those of the wild-type control during 8 days of cultivation. Thus, knockout of unrequired genes can reduce contamination with HCP in the production of recombinant therapeutic proteins.
CRISPR-U™ KO cell case 1
(a)The constructed sgRNA targeted a unique sequence in exon 4 of the Anxa2 gene. The amplicon sequence was analyzed in both directions.
CRISPR-U™ KO cell case 2
(b)The constructed sgRNA targeted the unique sequence in exon 2 of the Ctsd gene.The amplicon sequence was analyzed in both directions.
Anxa2 gene knockout cell line, construction of Ctsd gene knockout cell line
Characterization of protein expression from CHO knockout cell lines by SDS-PAGE and western blotting analysis.
(a) Anxa2-knockout cell lines.
(b) Ctsd-knockout cell lines. Cell culture supernatants and cell lysates were subjected to SDS-PAGE. Total proteins were detected by CBB staining. Western blotting analysis of each protein was performed using respective capture and detection antibodies.
The asterisk indicates the nonspecific band. The double asterisk indicates what is likely a fragment of cathepsin D.
Annexin A2 and cathepsin D were not detected in the cell culture supernatants or lysates of the corresponding knockout cell lines, suggesting successful exclusion of these impurities from host cells for therapeutic protein production. In addition, no truncated HCP was observed in the protein expression analysis of the Anxa2 and Ctsd knockout cell lines.
Reference:
Fukuda, N., Senga, Y., & Honda, S. (2019). Anxa2‐and Ctsd‐knockout CHO cell lines to diminish the risk of contamination with host cell proteins. Biotechnology progress, e2820.

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