DNA Repair Deficient Cell lines
DNA repair pathways maintain the integrity of the genome, reducing the onset of cancer, disease and aging. Conversely, the requirement for DNA repair and genome maintenance in response to radiation and genotoxic chemotherapeutics, positions DNA repair proteins as prime targets of anti-cancer drugs currently on clinical trials. Over 150 human proteins have been found with functional roles in DNA repair, that are categorized according to the main process they are involved in, including the classical DNA Repair pathways of Base Excision Repair (BER), Mismatch Excision Repair (MMR), Nucleotide Excision Repair (NER), Homologous Recombination (HR), and Non-Homologous End-Joining (NHEJ). Trevigen offers a broad panel of knockdown cell lines that will allow an in-depth study of the molecular etiology of genomic instability, providing useful tools to improve gene function analysis, and validation of targets during compound screenings.
Trevigen’s Knockdown Cell Lines are target specific LN428 (glioblastoma) shRNA lentivirus transduced cells. They are rigorously qualified and mycoplasma free. The percentage of knockdown levels range from 63 to 98% depending on the gene, as evaluated by RT-PCR. Expression of the shRNAs is maintained by puromycin selection.
All cell lines tested negative for mycoplasma and are provided under a material transfer agreement.
Knockdown Control (1)
The Knockdown Control Cell Line is a shRNA lentivirus transduced LN428 (glioblastoma) cell line, expressing a control non-targeting shRNA. Expression of the shRNA is maintained by puromycin selection.
The Knockdown Control is available upon request for any of the Knockdown Cell Line Pathways.
Base Excision Repair (19)
Base Excision Repair (BER) is an essential DNA repair pathway involved in the maintenance of genome stability by correcting DNA damage product of oxidation, deamination or alkylation. A DNA glycosylase initates the process recognizing and removing the damaged base, leaving empty or abasic site that will be later repaired by different proteins, preventing the occurrence of mutations (Krokan HE and Bjøra M, Cold Spring Harb Perspect Biol 2013;5:a012583).
The Knockdown Cell Lines offered include the following targeted genes: APE1, APE2, LIG3, MBD4, MPG, MutYh, NEIL1, NEIL2, NEIL3, NTHL1, OGG1, PARG, PARP1, PARP3, TDG, and XRCC1.
Homologous Recombination (7)
Homologous Recombination (HR) follows after Non-homologous end Joining (NHEJ) as the second major pathway for the repair of double DNA strand brakes (DSBs) in mammalian cells. This crucial DNA repair pathway only functions when homologous regions of DNA are available: late S and G2 phases of the cell cycle. Enzymes involved in HR use the homologous template to faithfully repair the DNA, and as a consequence, DSBs are repaired in a more accurately fashion (Bolderson E. et al., Clin Cancer Res 2009;15(20) October 15, 2009).
The Knockdown Cell Lines offered include the following targeted genes: BRCA1, NBS1 (NBN), RAD21, RAD50, RAD51C, SMC6L1, and XRCC3.
Mismatch Repair (7)
During DNA replication or after DNA damage, errors like mismatched bases or small insertions and deletions, are repaired by a system of enzymes and proteins called DNA mismatch repair (MMR). The MMR system, together with the DNA replication machinery, promote repair by a mechanism that involves the excision and resynthesis of DNA during or after DNA replication, increasing replication fidelity. Conversely, inactivation of genes part of MMR results in increased mutation rates. (Reyes, GX et al., 2014, Chromosoma, 124(4)).
The Knockdown Cell Lines offered include the following targeted genes: MLH1, MLH3, MSH2, MSH3, MSH5, MSH6, and PMS2.
Non-Homologous End Joining (5)
Non-homologous End Joining (NHEJ) repair mechanism involves, as its name indicates, the covalent union of broken DNA strands without the need of homology of sequence. NHEJ is more prone to errors than homology-based repair systems.
The Knockdown Cell Lines offered include the following targeted genes: SIRT1, SIRT2, SIRT3, PRKDC, and XRCC5.