DNA Polymerase theta (POLQ) Human Gene Knockout Kit (CRISPR)

CAT#: KN212566RB

POLQ - human gene knockout kit via CRISPR, HDR mediated

Product Images

Specifications

Product Data

• Format: 2 gRNA vectors, 1 RFP-BSD donor, 1 scramble control
• Donor DNA: RFP-BSD
• Symbol: DNA Polymerase theta
• Locus ID: 10721
• Components:

  KN212566G1, DNA Polymerase theta gRNA vector 1 in pCas-Guide CRISPR vector, Target Sequence: GATTCGTTCTCGGGAAGCGG

  KN212566G2, DNA Polymerase theta gRNA vector 2 in pCas-Guide CRISPR vector, Target Sequence: AAACGGCGGCGTTCAGAATC

  KN212566RBD, donor DNA containing left and right homologous arms and RFP-BSD functional cassette.

  Homologous arm and RFP-BSD sequences:

  pUC vector backbone in gray; Left arm sequence in blue; RFP-BSD in green; Right arm in violet

  AAGGCGAGTT ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT TCGGTCCTCC GATCGTTGTC AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG CAGCACTGCA TAATTCTCTT ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG AGTACTCAAC CAAGTCATTC TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG CGTCAATACG GGATAATACC GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA AACGTTCTTC GGGGCGAAAA CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT AACCCACTCG TGCACCCAAC TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT GAGCAAAAAC AGGAAGGCAA AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT GAATACTCAT ACTCTTCCTT TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT TTCCCCGAAA AGTGCCACCT GACGTCTAAG AAACCATTAT TATCATGACA TTAACCTATA AAAATAGGCG TATCACGAGG CCCTTTCGGG TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG TTGACGGTCA CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC ACCATAAAAT TGTAAACGTT AATATTTTGT TAAAATTCGC GTTAAATTTT TGTTAAATCA GCTCATTTTT TAACCAATAG GCCGAAATCG GCAAAATCCC TTATAAATCA AAAGAATAGC CCGAGATAGG GTTGAGTGTT GTTCCAGTTT GGAACAAGAG TCCACTATTA AAGAACGTGG ACTCCAACGT CAAAGGGCGA AAAACCGTCT ATCAGGGCGA TGGCCCACTA CGTGAACCAT CACCCAAATC AAGTTTTTTG GGGTCGAGGT GCCGTAAAGC ACTAAATCGG AACCCTAAAG GGAGCCCCCG ATTTAGAGCT TGACGGGGAA AGCCGGCGAA CGTGGCGAGA AAGGAAGGGA AGAAAGCGAA AGGAGCGGGC GCTAGGGCGC TGGCAAGTGT AGCGGTCACG CTGCGCGTAA CCACCACACC CGCCGCGCTT AATGCGCCGC TACAGGGCGC GTACTATGGT TGCTTTGACG TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA TACCGCATCA GGCGCCATTC GCCATTCAGG CTGCGCAACT GTTGGGAAGG GCGATCGGTG CGGGCCTCTT CGCTATTACG CCAGCTGGCG AAAGGGGGAT GTGCTGCAAG GCGATTAAGT TGGGTAACGC CAGGGTTTTC CCAGTCACGA CGTTGTAAAA CGACGGCCAG TGAATTGGAG GCTACAGTCA GTGGAGAGGA CTTTCACAGG CTGTCGCCGT GCTCATTTGA TAACTGCCCG TTATTCATGC GACACAATCC CCAATTCCTC ATTACATATT CCTCACAATG CATGCTGGCT AAAATGGAAA TTGTGTTCAG AAATAGCCCT GCCCTACCAT CATATACTAG GAGAAAGGAG GAAATTATGC TCTATTAGAA ACCCTAGGCT ATCTTGCATA ATGCTTCTCA ATCTGAAATA ATTTTTTACA GAATACCAAT ACGTTAAGAA GAGCTACTTC CCTGATCTAC CTATTATGGG CAGCCCATTT TATGAAGTTA GCATGGCCTT CCTATTCAAA CCAGAAATGG ACAAAAGGAG AAGGAAGGCT TTTAGGTCAG TATGGTTATA ACTGTAGATT CTGTATTCTT CCTCTTTAAT CTATCTCTCA TATTAACCAA CCAGGAGGTT GGAGGCCGGA AGTCTTAGAC GCCAGCGAGA GCCGCTCCTT CCCCACGAGT CTATGGCTTT CGGGTTACTT AAGCCACGGA GAACTCTATG GTTCCGGGGC GGGCCGGGAG GTTTGAGTTT GAAGACTGGC GGGAAGATGT CCGCAGCTGT TGCCAGGCCA GGGTTCTCCC GAGAGGGAGG ACGCTGGGAC TGTGGCTTGC CCTGATCGGC CGAGAAGAGT TTGCCCCTGA AGGCCGCAGC TGCAGGTGCT CCAGGAACCC GGGAGGCCGC GGGCAACGGG GGCAGCTCCG CCGGGCCATG GGTCGGGAGG TGTCCTCACC CCACGGGACT TGCTTCGTTG CATACCCCAT TGGGCTTCTA CTCCCCTTCT CTGTGACTGG GGCCTGGTGT GGCAGGTCAC TTTCTGTTCA GACATACGCG TGTTTTCAGT GATCGCCTGG TATTAATGTT CACGTGGCTT CTGGATCTTT CGACCTTTAG AGAAACAACT TCCTAGTTAG CTTCATCCCA TCATCTGTTG GGCGGCTCGT TCCTGGTAGT ACCCAGTACT CCTTACAGAC CCTTAAGACA GGATTGTTGG GCCTGGAGTA CGGCACAGGC TGTTCGGCGT CCAGGGGAAG GCTGGAAGGG ATTCGTAATG AGATTGGTTG GGTGTCCCGT GGATGTCGAC CCAGTTTTTT AAAATCGTTA GTTGTGCAGT CGTCATGGAA CTAAAACTTA CTATAAATAG AAACTCACCT AACTTAGCTT TGTAATGCTT GCATATTGCC CTCCTCCCCC AACTACACCC GTAATTCCAA CACTCTTTAT CTACTGTGTA TTGCATCACT CTCGCCGGTT GGACTTTAGA TCAGAAGGGA TCTTGCTGCC GCCCGAAAGA GGAAGGGCTG GAAGAGGAAG GAGCTTGGCG TAATCATGGT CATAGCTGTT TCCTGTGTGA AATTGTTATC CGCTCACAAT TCCACACAAC ATACGAGCCG GAAGCATAAA GTGTAAAGCC TGGGGTGCCT AATGAGTGAG CTAACTCACA TTAATTGCGT TGCGCTCACT GCCCGCTTTC CAGTCGGGAA ACCTGTCGTG CCAGCTGCAT TAATGAATCG GCCAACGCGC GGGGAGAGGC GGTTTGCGTA TTGGGCGCTC TTCCGCTTCC TCGCTCACTG ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC GAGCGGTATC AGCTCACTCA AAGGCGGTAA TACGGTTATC CACAGAATCA GGGGATAACG CAGGAAAGAA CATGTGAGCA AAAGGCCAGC AAAAGGCCAG GAACCGTAAA AAGGCCGCGT TGCTGGCGTT TTTCCATAGG CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA GTCAGAGGTG GCGAAACCCG ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT CCCTCGTGCG CTCTCCTGTT CCGACCCTGC CGCTTACCGG ATACCTGTCC GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT TCTCATAGCT CACGCTGTAG GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC TGTGTGCACG AACCCCCCGT TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG CAGCCACTGG TAACAGGATT AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA AGTGGTGGCC TAACTACGGC TACACTAGAA GAACAGTATT TGGTATCTGC GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA GCTCTTGATC CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC AGATTACGCG CAGAAAAAAA GGATCTCAAG AAGATCCTTT GATCTTTTCT ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT CATGAGATTA TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT GAAGTTTTAA ATCAATCTAA AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT TAATCAGTGA GGCACCTATC TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC TCCCCGTCGT GTAGATAACT ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA TGATACCGCG AGAACCACGC TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG GAAGGGCCGA GCGCAGAAGT GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT GTTGCCGGGA AGCTAGAGTA AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA TTGCTACAGG CATCGTGGTG TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT CCCAACGATC

  GE100003, scramble sequence in pCas-Guide vector

• Disclaimer: These products are manufactured and supplied by OriGene under license from ERS. The kit is designed based on the best knowledge of CRISPR technology. The system has been functionally validated for knocking-in the cassette downstream the native promoter. The efficiency of the knock-out varies due to the nature of the biology and the complexity of the experimental process.

Reference Data

• RefSeq: NM_006596, NM_199420
• UniProt ID: O75417
• Synonyms: DKFZp781A0112; DNA polymerase eta; DNA polymerase theta; POLH; POLH, PRO0327, DKFZp781A0112; polymerase (DNA directed), theta; PRO0327
• Summary: DNA polymerase that promotes microhomology-mediated end-joining (MMEJ), an alternative non-homologous end-joining (NHEJ) machinery triggered in response to double-strand breaks in DNA (PubMed:25642963, PubMed:25643323). MMEJ is an error-prone repair pathway that produces deletions of sequences from the strand being repaired and promotes genomic rearrangements, such as telomere fusions, some of them leading to cellular transformation (PubMed:25642963, PubMed:25643323). POLQ acts as an inhibitor of homology-recombination repair (HR) pathway by limiting RAD51 accumulation at resected ends (PubMed:25642963). POLQ-mediated MMEJ may be required to promote the survival of cells with a compromised HR repair pathway, thereby preventing genomic havoc by resolving unrepaired lesions (By similarity). The polymerase acts by binding directly the 2 ends of resected double-strand breaks, allowing microhomologous sequences in the overhangs to form base pairs. It then extends each strand from the base-paired region using the opposing overhang as a template. Requires partially resected DNA containing 2 to 6 base pairs of microhomology to perform MMEJ (PubMed:25643323). The polymerase activity is highly promiscuous: unlike most polymerases, promotes extension of ssDNA and partial ssDNA (pssDNA) substrates (PubMed:18503084, PubMed:21050863, PubMed:22135286). Also exhibits low-fidelity DNA synthesis, translesion synthesis and lyase activity, and it is implicated in interstrand-cross-link repair, base excision repair and DNA end-joining (PubMed:14576298, PubMed:18503084, PubMed:19188258, PubMed:24648516). Involved in somatic hypermutation of immunoglobulin genes, a process that requires the activity of DNA polymerases to ultimately introduce mutations at both A/T and C/G base pairs (By similarity).[UniProtKB/Swiss-Prot Function]