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 typeVarious types of cells including tumor cell lines, regular cell lines, IPS/ES cell lines
Service typePoint Mutation / Reporter Gene Knock-in
DeliverablesCell pool/Single-cell clone
Turnaround/Price8 weeks，as low as $6480   Contact Us
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(293T)
Human prostate carcinoma cell line (22RV1)
Madin-Darby Canine Kidney Cell Line (MDCK)
Distal nephron cell line (Distal nephron cell line（JU4s）)

Blood and Lymphatic System

Mouse Macrophage Cell Line (RAW264.7)
Porcine Alveolar Macrophage Cell Line (3D4/21)
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

Respiratory System

Human Lung Cancer Cell Line (A549)
Human Lung Cancer Cell Line (Calu-1)
Human Lung Squamous Carcinoma Cell Line (NCI-H520)
Lewis lung carcinoma (LLC)
Human Lung Cancer Cell Line (SK-MES-1)
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 Epithelial Cell Line (HBL-100)
Human Breast Cancer Cell Line (MCF7)
Mouse Breast Cancer Cell Line (4T1)
Human Pancreatic Carcinoma Cell Line (PANC-1)
Human Breast Cancer Cell Line (JIMT-1)
Human breast cancer cell line (MCF-7)
Human Breast Adenocarcinoma Cell Line (SK-BR-3)
Human Metastatic Pancreatic Adenocarcinoma Cell Line (AsPC-1)
Mouse Pancreatic Cancer Cell Line (Pan02)
Murine Breast Cancer Cell Line (E0771)
Human Pancreatic Carcinoma Cell Line (MIA PaCa-2)
Human prostate cancer cell line (PC3)
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-47D)

Circulatory System

Rat Cardiac Myocytes (H9C2)
Mouse Cardiac Muscle Cell Line (HL-1)
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 Cell Line (L-02)
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 Hepatocarcinoma Cell Line (SMMC-7721)
Mouse Hepatoma Cell Line (H22)
Mouse Hepatoma Cell Line (Hepa 1-6)
Human hepatobiliary cancer cell line (RBE)
Human Gastric Cancer Cell Line (HGC-27)
Human Gastric Cancer Cell Line (SGC-7901)
Human Gastric Cancer Cell Line (MGC80-3)
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)
Human Renal Cell Carcinoma Cell Line (ACHN)

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)
Human Epidermoid Carcinoma Cell Line (A431)
Mouse myeloma cell line (Sp2/0-Ag14)
Mouse Skin Cancer Cell Line (SCC7)
Immortalized Ameloblastoma Cells (hTERT-AM)

Ocular, Otolaryngologic and Oral System

Rat Muller Cell Line (rmc-1)
Human Nasopharyngeal Carcinoma Cell Line (NPC-43)
Human Nasopharyngeal Carcinoma Cell Line (cne2z)
Human Nasopharyngeal Carcinoma Cell Line (CNE-1)

Brain and Nervous System

Human Neuroblastoma Cell Line (SK-N-SH)
Human Neuroblastoma Cell Line (SH-SY5Y)
Mouse neuroblastoma cell line (Neuro-2a)
Human Glioblastoma Cell Line (U251)
Rat Glioblastoma Cell Line (C6)
Mouse Glioblastoma Cell Line (GL261)
Mouse Microglial Cell Line (BV2)
Human glioblastoma cell line (U-87 MG)
Mouse Anterior Parietal Bone Cell Line (MC3T3-E1 Subclone 14)
Human Microvascular Endothelial Cell Line (hCMECD3)
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)
Human Choriocarcinoma Cell Line (BeWo)
Human Ovarian Cancer Cell Line (CAOV3)
Human Trophoblast Cell Line (HTR-8/SVneo)
Mouse Pituitary Gonadotrope Cell Line (Lbetat2)
Mouse Testicular Stromal Cell Line (TM3)

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
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.

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