Knockin Cell Lines

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CRISPR-U™ Cell Line
CRISPR-U™ (based on CRISPR/Cas9 technology), 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) in vitro and in vivo. With CRISPR-U, Ubigene has successfully edit genes on more than 100 cell lines.
CRISPR/Cas9 recognizes the target sequence with gRNA, and guide Cas9 endonuclease to cut the upstream of PAM, resulting in the double-strand break (DSB) of the target site DNA. To repair the DSB, the cell uses its own DNA repair mechanism to add or delete or replace pieces of DNA sequences via Homology Directed Repair (HDR) or Non-Homologous End Joining (NHEJ).Close
CRISPR-U™ (based on CRISPR/Cas9 technology), 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
Gene-editing Cases : 100 Types of Cell Lines

Gene-editing Cases:100 Types of Cell LinesClick Here

Urinary System

Human bladder carcinoma cell line (TCCSUP)
Human bladder carcinoma cell line (5637)
Human Bladder Transitional Cell Carcinoma Cell Line (T24)
Human Embryonic Kidney Cell Line(HEK293)
Human Embryonic Kidney Cell Line(derived from 293)
Human Embryonic Kidney Cell Line(293T)
Human prostate carcinoma cell line (22RV1)
Madin-Darby Canine Kidney Cell Line MDCK(NBL-2)

Blood and Lymphatic System

Mouse Macrophage Cell Line (RAW264.7)
Human Monocytic Cell Line (THP-1)
Rat Basophil Leukemia Cell Line (RBL-2H3)
Human leukemia cell line (HL-60)
Human T lymphocyte cell line (Jurkat, Clone E6-1)
Human myelogenous leukemia cell line (K-562)
Human caucasian histiocytic lymphoma cell line (U-937)
Human Acute Non-B Non-T Lymphocytic Leukemia Cell Line (Reh)
Human B Cell Lymphoma Cancer Cell Line U2932
Human B Cell Lymphoma Cancer Cell Line OCI-LY3

Respiratory System

Human Lung Cancer Cell Line (A549)
Human Lung Cancer Cell Line (Calu-1)
Lewis lung carcinoma (LLC)
Human Lung Squamous Cell Carcinoma Cell Line NCI-H226
Human Non-small Cell Lung Carcinoma Cell Line (NCI-H1299)
Human Non-small Cell Lung Carcinoma Cell Line (HCC827)
Human Small Cell Lung Cancer Cell Line H69AR
Human Bronchial Epithelial Cell Line (BEAS-2B)
Human Bronchial Epithelial Cell Line (16HBE)
Human hypopharyngeal carcinoma cell line (FaDu)
Chinese hamster lung cells (V79)

Endocrine System

Human Breast Cancer Cell Line (MDA-MB-231)
Human Breast Cancer Cell Line (MDA-MB-468)
Human Breast Cancer Cell Line (MDA-MB-453)
Human Breast Cancer Cell Line (MDA-MB-436)
Human Breast Cancer Cell Line (ZR-75-1)
Human Breast Cell Line (MCF10A)
Rat Breast Cancer Cell Line (4T1)
Human Pancreatic Carcinoma Cell Line (PANC-1)
Mouse Pancreatic Carcinoma Cell Line (Capan-2)
Human breast cancer cell line (MCF-7)
Human Ductal Carcinoma Cell Line (BT-474)
Human Breast Adenocarcinoma Cell Line (SK-BR-3)
Human Metastatic Pancreatic Adenocarcinoma Cell Line (AsPC-1)
Human Pancreatic Carcinoma Cell Line (MIA PaCa-2)
Human prostate cancer cell line (PC3)
Human prostate cancer cell line (LNCaP)
Human Prostate Cancer Cell Line (VCaP)
Mouse Acinar Pancreatic Cell Line (266-6)
Human pancreatic cancer cell line (BxPC-3)
Human Breast Adenocarcinoma Cell Line (T-4D)

Circulatory System

Rat Cardiac Myocytes (H9C2)
Human Coronary Artery Endothelial Cell line (HCAEC)
Mouse Myoblast Cell Line (C2C12)

Digestive System

Human Colon Cancer Cell Line (HCT116)
Human Colon Cancer Cell Line (SW480)
Human Colon Cancer Cell Line (SW620)
Human Colon Cancer Cell Line (HT-29)
Human Colon Cancer Cell Line (LoVo)
Human colon carcinoma cell line (RKO)
Human caucasian colon adenocarcinoma cell line (COLO 205)
Murine Colorectal Carcinoma Cell Line (MC38)
Murine colorectal carcinoma cell line (CT26.WT)
Human colon adenocarcinoma cell line (DLD-1)
Human colorectal adenocarcinoma cell line (NCI-H716)
Human colorectal adenocarcinoma cell line (Caco-2)
Human colon carcinoma cell line (T84)
Human liver cancer cell line (Hep G2)
Human Hepatoma Cell Line (Hep 3B)
Human hepatocellular carcinoma cell line (HuH-7)
Human Hepatocellular Carcinoma Cell Line (SNU-387)
Human hepatobiliary cancer cell line (RBE)
Human Gastric Cancer Cell Line (HGC-27)
Human gastric cancer cell line (AGS)
Human Esophageal Squamous Carcinoma Cell Line (KYSE-150)
Human Esophageal Squamous Carcinoma Cell Line (KYSE-30)
Human renal cell carcinoma cell line (786-O)
African green monkey kidney cell (Vero)

Skeleton, Articulus, Soft Tissue, Derma System

Human Osteosarcoma Cell Line (MG63)
Human bone osteosarcoma epithelial cell line (U-2 OS)
Human fibrosarcoma cell line (HT1080)
Human malignant melanoma cell line (A-375)
Human Melanoma Cell Line (M14)
Murine melanoma cell line (B16-F10)
Mouse myeloma cell line (Sp2/0-Ag14)

Ocular, Otolaryngologic and Oral System

Rat Muller Cell Line (rmc-1)
Human Nasopharyngeal Carcinoma Cell Line (C666-1)
Human Nasopharyngeal Carcinoma Cell Line (cne2z)
Human Nasopharyngeal Carcinoma Cell Line (CNE-1)

Brain and Nervous System

Human Neuroblastoma Cell Line (SK-N-SH)
Mouse neuroblastoma cell line (Neuro-2a)
Human Glioblastoma Cell Line (U251)
Rat Glioblastoma Cell Line (C6)
Mouse Glioblastoma Cell Line (GL261)
Human glioblastoma cell line (U-87 MG)
Mouse Anterior Parietal Bone Cell Line (MC3T3-E1 Subclone 14)
Immortalize Human Microvascular Endothelial Cell Line (hCMEC/D3)
Mouse Hippocampal Neuron Cell Line (HT22)

Reproductive System

Human Cervical Carcinoma Cell Line (HeLa)
Human Cervical Carcinoma Cell Line (Hela 229)
Human Cervical Squamous Cell Line (SiHa)
Mouse Embryonic Fibroblasts (NIH/3T3)
Chinese Hamster Ovary Cell Line (CHO-K1)
Human Ovarian Cancer Cell Line (SK-OV-3)
Human Ovarian Cancer Cell Line (OVCAR3)
Mouse Testicular Stromal Cell Line (TM3)

Technical advantage
CRISPR-U:10X efficiency gene-editing

Exclusive innovation, 10 times more efficient in gene-editing.

100 types of knockin cell line

Successfully edit genes on more than 100 types of cell lines.

KO/PM/ KI

Easily generate knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo.

100% mutation guarantee

CRISPR-U™ offers a 100% mutation guarantee. No mutation, no charge!

Gene Knockin Cell Line
The cells would be co-transfected with gRNA, Cas9 and donor vector by electroporation, and then screened. After drug screening, single clones would be generated. Positive clones would be validated by sequencing. Cells knockin with reporter genes can also be selected by observing the fluorescence protein.
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 flow
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.

B

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.