knockin cell line

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Knockin Cell Line

The gRNA vector (containing Cas9) of the CRISPR/Cas9 system and the Donor carrying target mutation or target gene sequences will be co-transfected into cells by electroporation. Then the gRNA and Cas9 complex will cause a double-strand break (DSB) on the target site of DNA and the cells will undergo homologous recombination repair (HDR) using Donor as a template to recombine target mutations to the target sites. After antibiotic screening, single-cell clones will be generated. Positive clones that are successfully introduced the target mutations be validated by target site amplification and sequencing. Final deliverables will be the positive cell clones, related data and project reports.
Knockin Cell Service Details
Cell type Various types of cells including tumor cell lines, regular cell lines, IPS/ES cell lines
Service type Point Mutation / Reporter Gene Knock-in
Deliverables Cell pool/Single-cell clone
Turnaround/Price 8 weeks,as low as $6480   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

Knockin Strategies
Protein fusion
Guide RNA and Cas9 complex cause a double-strand break (DSB) on the target site of DNA. The donor vector carrying knockin sequence is the template for homologous recombination repair (HDR), and it recombines to the target site.
Protein fusion Knockin cell
Replacement of specific locus:
knockin cell specific locus
Work Flow and Validation
Knockin Cell Line Work Flow and ValidatioKnockin Cell Line Work Flow and Validatio
Case Study
T cells carrying chimeric antigen receptors (CARs) can mediate tumor rejection, which is an effective way to cure B cell malignant tumors. By using CRISPR/Cas9 technology, TRAC gene was replaced by CD19 specific CAR sequence.CAR expression in human peripheral blood T cells was found, and the immune response ability of T cells was enhanced.
A

CRISPR/Cas9 mediated CAR gene integrated into TARC site.
1928z is a CD19 specific CAR sequence.

CRISPR/Cas9 mediated CAR gene integrated into TARC site.1928z is a CD19 specific CAR sequence.
B

TCR/CAR flow plots.4 days after co-transfection of
Cas9, gRNA and donor vector, CAR proteins were detected.

TCR/CAR flow plots.4 days after co-transfection of Cas9, gRNA and donor vector, CAR proteins were detected.
Reference:
Eyquem, J., Mansilla-Soto, J., Giavridis, T., van der Stegen, S. J., Hamieh, M., Cunanan, K. M., ... & Sadelain, M. (2017). Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection. Nature, 543(7643), 113.

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