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

CAT#: KN212566

POLQ - human gene knockout kit via CRISPR, HDR mediated

Product Images

Specifications

Product Data

• Format: 2 gRNA vectors, 1 GFP-puro donor, 1 scramble control
• Donor DNA: GFP-puro
• 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

  KN212566D, donor DNA containing left and right homologous arms and GFP-puro functional cassette.

  Homologous arm and GFP-puro sequences:

  pUC vector backbone in gray; Left arm sequence in blue; GFP-puro 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 TTGCCACTAG CATGGAGAGC GACGAGAGCG GCCTGCCCGC CATGGAGATC GAGTGCCGCA TCACCGGCAC CCTGAACGGC GTGGAGTTCG AGCTGGTGGG CGGCGGAGAG GGCACCCCCG AGCAGGGCCG CATGACCAAC AAGATGAAGA GCACCAAAGG CGCCCTGACC TTCAGCCCCT ACCTGCTGAG CCACGTGATG GGCTACGGCT TCTACCACTT CGGCACCTAC CCCAGCGGCT ACGAGAACCC CTTCCTGCAC GCCATCAACA ACGGCGGCTA CACCAACACC CGCATCGAGA AGTACGAGGA CGGCGGCGTG CTGCACGTGA GCTTCAGCTA CCGCTACGAG GCCGGCCGCG TGATCGGCGA CTTCAAGGTG ATGGGCACCG GCTTCCCCGA GGACAGCGTG ATCTTCACCG ACAAGATCAT CCGCAGCAAC GCCACCGTGG AGCACCTGCA CCCCATGGGC GATAACGATC TGGATGGCAG CTTCACCCGC ACCTTCAGCC TGCGCGACGG CGGCTACTAC AGCTCCGTGG TGGACAGCCA CATGCACTTC AAGAGCGCCA TCCACCCCAG CATCCTGCAG AACGGGGGCC CCATGTTCGC CTTCCGCCGC GTGGAGGAGG ATCACAGCAA CACCGAGCTG GGCATCGTGG AGTACCAGCA CGCCTTCAAG ACCCCGGATG CAGATGCCGG TGAAGAAAGA GTTTAAGAAT TCCGATCATA TTCAATAACC CTTAATATAA CTTCGTATAA TGTATGCTAT ACGAAGTTAT TAGGTCTGAA GAGGAGTTTA CGTCCAGCCA AGCTTAGGAT CTCGACCTCG AAATTCTACC GGGTAGGGGA GGCGCTTTTC CCAAGGCAGT CTGGAGCATG CGCTTTAGCA GCCCCGCTGG GCACTTGGCG CTACACAAGT GGCCTCTGGC CTCGCACACA TTCCACATCC ACCGGTAGGC GCCAACCGAC TCCGTTCTTT GGTGGCCCCT TCGCGCCACC TTCTACTCCT CCCCTAGTCA GGAAGTTCCC CCCCGCCCCG CAGCTCGCGT CGTGCAGGAC GTGACAAATG GAAGTAGCAC GTCTCACTAG TCTCGTGCAG ATGGACAGCA CCGCTGAGCA ATGGAAGCGG GTAGGCCTTT GGGGCAGCGG CCAATAGCAG CTTTGCTCCT TCGCTTTCTG GGCTCAGAGG CTGGGAAGGG GTGGGTCCGG GGGCGGGCTC AGGGGCGGGC TCAGGGGCGG GGCGGGCGCC CGAAGGTCCT CCGGAGGCCC GGCATTCTGC ACGCTTCAAA AGCGCACGTC TGCCGCGCTG TTCTCCTCTT CCTCATCTCC GGGCCTTTCG ACCTGCATCC ATCTAGATCT CGAGCAGCTG AAGCTTACCA TGACCGAGTA CAAGCCCACG GTGCGCCTCG CCACCCGCGA CGACGTCCCC AGGGCCGTAC GCACCCTCGC CGCCGCGTTC GCCGACTACC CCGCCACGCG CCACACCGTC GATCCGGACC GCCACATCGA GCGGGTCACC GAGCTGCAAG AACTCTTCCT CACGCGCGTC GGGCTCGACA TCGGCAAGGT GTGGGTCGCG GACGACGGCG CCGCGGTGGC GGTCTGGACC ACGCCGGAGA GCGTCGAAGC GGGGGCGGTG TTCGCCGAGA TCGGCCCGCG CATGGCCGAG TTGAGCGGTT CCCGGCTGGC CGCGCAGCAA CAGATGGAAG GCCTCCTGGC GCCGCACCGG CCCAAGGAGC CCGCGTGGTT CCTGGCCACC GTCGGCGTCT CGCCCGACCA CCAGGGCAAG GGTCTGGGCA GCGCCGTCGT GCTCCCCGGA GTGGAGGCGG CCGAGCGCGC CGGGGTGCCC GCCTTCCTGG AGACCTCCGC GCCCCACAAC CTCCCCTTCT ACGAGCGGCT CGGCTTCACC GTCACCGCCG ACGTCGAGGT GCCCGAAGGA CCGCGCACCT GGTGCATGAC CCGCAAGCCC GGTGCCTGAC GCCCGCCCCA CGACCCGCAG CGCCCGACCG AAAGGAGCGC ACGACCCCAT GCATCGATGA TATCAGATCC CCGGGATGCA GAAATTGATG ATCTATTAAA CAATAAAGAT GTCCACTAAA ATGGAAGTTT TTCCTGTCAT ACTTTGTTAA GAAGGGTGAG AACAGAGTAC CTACATTTTG AATGGAAGGA TTGGAGCTAC GGGGGTGGGG GTGGGGTGGG ATTAGATAAA TGCCTGCTCT TTACTGAAGG CTCTTTACTA TTGCTTTATG ATAATGTTTC ATAGTTGGAT ATCATAATTT AAACAAGCAA AACCAAATTA AGGGCCAGCT CATTCCTCCC ACTCATGATC TATAGATCTA TAGATCTCTC GTGGGATCAT TGTTTTTCTC TTGATTCCCA CTTTGTGGTT CTAAGTACTG TGGTTTCCAA ATGTGTCAGT TTCATAGCCT GAAGAACGAG ATCAGCAGCC TCTGTTCCAC ATACACTTCA TTCTCAGTAT TGTTTTGCCA AGTTCTAATT CCATCAGAAG CTGGTCGAGA TCCGGAACCC TTAATATAAC TTCGTATAAT GTATGCTATA CGAAGTTATT AGGTCCCTCG AAGAGGTTCA CTAGGCGCGC CCCTGAAGGC CGCAGCTGCA GGTGCTCCAG GAACCCGGGA GGCCGCGGGC AACGGGGGCA GCTCCGCCGG GCCATGGGTC GGGAGGTGTC CTCACCCCAC GGGACTTGCT TCGTTGCATA CCCCATTGGG CTTCTACTCC CCTTCTCTGT GACTGGGGCC TGGTGTGGCA GGTCACTTTC TGTTCAGACA TACGCGTGTT TTCAGTGATC GCCTGGTATT AATGTTCACG TGGCTTCTGG ATCTTTCGAC CTTTAGAGAA ACAACTTCCT AGTTAGCTTC ATCCCATCAT CTGTTGGGCG GCTCGTTCCT GGTAGTACCC AGTACTCCTT ACAGACCCTT AAGACAGGAT TGTTGGGCCT GGAGTACGGC ACAGGCTGTT CGGCGTCCAG GGGAAGGCTG GAAGGGATTC GTAATGAGAT TGGTTGGGTG TCCCGTGGAT GTCGACCCAG TTTTTTAAAA TCGTTAGTTG TGCAGTCGTC ATGGAACTAA AACTTACTAT AAATAGAAAC TCACCTAACT TAGCTTTGTA ATGCTTGCAT ATTGCCCTCC TCCCCCAACT ACACCCGTAA TTCCAACACT CTTTATCTAC TGTGTATTGC ATCACTCTCG CCGGTTGGAC TTTAGATCAG AAGGGATCTT GCTGCCGCCC GAAAGAGGAA GGGCTGGAAG AGGAAGGAGC TTGGCGTAAT CATGGTCATA GCTGTTTCCT GTGTGAAATT GTTATCCGCT CACAATTCCA CACAACATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG GTGCCTAATG AGTGAGCTAA CTCACATTAA TTGCGTTGCG CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAA CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC GTGGTGTCAC GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATC

  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]