2016-5-23 · This plasmid is referred to as pX260-MYC 3′ TBE1. The oligonucleotide sequences corresponding to the guide RNA are listed in Table S1. To design the MYC 3′ WRE-specific CRISPR reporter plasmid we employed a stepwise approach using standard PCR and sub-cloning procedures. The pEGFP-N1 plasmid (Clontech 6085-1 Mountain View CA USA
In order to target a given site the plasmid can be digested using BbsI and a pair of annealed oligos (design is indicated below) can be cloned into the CRISPR array. The oligos is designed based on the target site sequence (30bp) and needs to be flanked on the 3′ end by a 3bp NGG PAM sequence. Genbank Map of Backbone Plasmid PX260 (rev
2015-6-5 · Figure S1. CRISPR-mediated targeting of the OCT4 locus through drug selection. (A) The vector map of the px260 plasmid for expressing Cas9 and crRNA/tracrRNA. (B) Four crRNAs were designed to target the stop codon of the OCT4 locus.Each crRNA was cloned into the px260 vector and transfected into 293T cells.
pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro was a gift from Feng Zhang (Addgene plasmid # 42229) Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong L Ran FA Cox D Lin S Barretto R Habib N Hsu
2016-3-4 · For gRNA expression screening specific reverse primer (pX260-crRNA-3′/R Table I.3 in S1 Experimental Procedures) was used in RT reaction followed by standard PCR using target A or B
2020-12-14 · 1. pX260 (or pX334) S. pyogenes Cas9 (or Cas9 D10A nickase) CRISPR RNA array tracrRNA This plasmid contains three expression cassettes. In order to target a given site the plasmid can be digested using BbsI and a pair of annealed oligos (design is indicated below) can be cloned into the CRISPR array.
Plasmid pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro from Dr. Feng Zhang s lab contains the insert humanized S. pyogenes Cas9 and is published in Science. 2013 Jan 3. This plasmid is available through Addgene.
2021-1-1 · The pX260 system uses gRNA that carries Cas9 tracrRNA and CRISPR RNA array from S. pyogenes. Generally after digestion with restriction enzyme the plasmids are annealed with oligonucleotides that are designed for specific target sites . Despite of its usage this plasmid based system encounters quite a lot of defy in their mechanism.
2021-7-2 · The first delivery pattern is a plasmid such as pX260 and pX335 which encodes both sgRNA and Cas9. The second delivery pattern is a mixture of sgRNA and Cas9 mRNA. The third delivery pattern is Cas9/sgRNA ribonucleoprotein (RNP). 28 29 Various advantages and shortcomings lie in each delivery pattern for clinical translation.
pX260. Zhang lab vector for expressing Cas9 a tracrRNA and a guide sequence. Also known as pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro. To see this sequence with restriction sites features and translations please download
2020-12-14 · 1. pX260 (or pX334) S. pyogenes Cas9 (or Cas9 D10A nickase) CRISPR RNA array tracrRNA This plasmid contains three expression cassettes. In order to target a given site the plasmid can be digested using BbsI and a pair of annealed oligos (design is indicated below) can be cloned into the CRISPR array.
2017-1-22 · To construct pSicoR-sgRNAs-mCherry-Cas9 vector the puromycin fragment in pSico-EF1-mCh-Puro (Addgene Plasmid 31845) was replaced with the NLS-Cas9-NLS fragment from pX260 (Addgene Plasmid 42229) .Then the gRNA scaffold with the cloning site containing two BsmBI sites was inserted downstream of the U6 promoter.The schematic of pSicoR-sgRNA-mCherry-Cas9 vector was
2020-7-7 · PX260 This plasmid separately encodes a human codon-optimized SpCas9 a tracrRNA and customizable crRNA. #42230 PX330 A human codon-optimized SpCas9 and chimeric guide RNA expression plasmid. #48138 PX458 GFP Cas9 from S. pyogenes
2020-8-14 · px260 (Addgene plasmid no. 42229) pCMV-BE3 (Addgene plasmid no. 73021) pCAG-Cre (Yang lab no. ZP156) FastDigest BbsI (Thermo Fisher Scientific . no. FD1014)
1982-2-1 · pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro was a gift from Feng Zhang (Addgene plasmid # 42229) Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong L Ran FA Cox D Lin S Barretto R Habib N Hsu
Show Static Map. hCas9 9553 bp. Cas9. 748 .. 4851 = 4104 bp. 1368 amino acids = 158.4 kDa. Product Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system. generates RNA-guided double strand breaks in DNA.
px260. In Category. Educational Resources. Filter By. Expression. Mammalian Expression (1) Vector Types. CRISPR (1) Plasmid Type.
This plasmid is referred to as pX260-MYC 3′ TBE1. The oligonucleotide sequences corresponding to the guide RNA are listed in Table S1. To design the MYC 3′ WRE-specific CRISPR reporter plasmid we employed a stepwise approach using standard PCR and sub-cloning procedures. The pEGFP-N1 plasmid (Clontech 6085-1 Mountain View CA USA
2014-7-18 · Plasmid Preparation. DNA segment expressing long-term repeats (LTR)-A or LTR-B for precrRNA was cloned into the pX260 vector that contains the puromycin selection gene (Addgene plasmid #42229). DNA segment expressing LTR-C or LTR-D for the chimeric crRNA-tracrRNA was cloned into the pX330 vector (Addgene plasmid #42230). Both vectors contain
Plasmid pX330-U6-Chimeric_BB-CBh-hSpCas9 from Dr. Feng Zhang s lab contains the insert humanized S. pyogenes Cas9 and is published in Science. 2013 Jan 3. This plasmid is available through Addgene. More than 20 requests
pX260 pX330 pX330A-1x2 pX330S-2 pX334 pX335 PX551 PX552 pX600 pX601 pX602 PX851 PX852 PX853 PX854 PX855 PX856 pY010 (pcDNA3.1-hAsCpf1) pY016 (pcDNA3.1-hLbCpf1) sgRNA(MS2) cloning backbone SP-Cas9 SP-dCas9-VPR VP12
2021-4-27 · 2.5 mg pX260 plasmid and 2.5 mg donor plasmid with GFP by electroporator (Nepa Gene Chiba Japan). Transfected single cells were recovered in NutriStem hPSC XF Medium (Biological Industries Beit-Haemek Israel) and seeded on Matrigel-coated plates. Four days after electroporation the culture medium was changed back to mTeSR1 medium and
2020-8-14 · px260 (Addgene plasmid no. 42229) pCMV-BE3 (Addgene plasmid no. 73021) pCAG-Cre (Yang lab no. ZP156) FastDigest BbsI (Thermo Fisher Scientific . no. FD1014)
2017-11-28 · The pX260 system also known as the pX334 system contains three cassettes including a CRISPR RNA array tracrRNA and S. pyogenes Cas9 (or the Cas9 D10A nickase). The plasmid is digested with a restriction enzyme and then ligated with an annealed oligonucleotide that is designed for a specific targeting site.
1982-2-1 · pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro was a gift from Feng Zhang (Addgene plasmid # 42229) Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong L Ran FA Cox D Lin S Barretto R Habib N Hsu
pX260 #42229 pX330 #42230 pX335 #42335 pX458 #48138 pX458M EZ-GuideXH pX459M EZ-GuideXH EZ-GuideXH pX461 #48140 pX552 #60958 pX601 #61591 pX602 #61593 LentiCRISPR V2 #52961 pLen
2020-12-14 · 1. Digest 1ug of plasmid with . Bbs. I for. 30 min at 37°C 1 ug Plasmid . 1 ul FastDigest . Bbs. I (Fermentas) 1 ul FastAP (Fermentas) 2 ul 10X FastDigest Buffer . X ul ddH. 2 O 20 ul total 2. Gel purify digested plasmid using. QIAquick Gel Extraction Kit and elute in EB. 3. Phosphorylate and anneal each pair of oligos 1 ul oligo 1 (100uM)
2017-9-11 · For instance in the pX260 or pX330 system Cas9 protein and sgRNA were expressed form the same plasmid. The pX260 system also known as the pX334 system contains three cassettes including a CRISPR RNA array tracrRNA and S. pyogenes Cas9 (or the Cas9 D10A nickase).
2017-11-28 · The pX260 system also known as the pX334 system contains three cassettes including a CRISPR RNA array tracrRNA and S. pyogenes Cas9 (or the Cas9 D10A nickase). The plasmid is digested with a restriction enzyme and then ligated with an annealed oligonucleotide that is designed for a specific targeting site.
2014-9-25 · Linearize plasmid PX260 (Addgene plasmid #42229) using the NcoI restriction site which is located immediately upstream of the initial codon of the NLS-hSpCas9-NLS expression cassette. Ligate the
2019-7-16 · PX260 This plasmid separately encodes a human codon-optimized SpCas9 a tracrRNA and customizable crRNA. #42230 PX330 A human codon-optimized SpCas9 and chimeric guide RNA expression plasmid. #48138 PX458 GFP Cas9 from S. pyogenes
2014-8-7 · Puro-Cas9 donor (Figure S1E) was constructed by replacing EGFP in the TRE-TIGHT-EGFP-BW plasmid (Addgene plasmid 22077) (Hockemeyer et al. 2011) with the human codon-optimized Streptococcus pyogenes Cas9 cDNA amplified by PCR from pX260 (Addgene plasmid
pX260 #42229 pX330 #42230 pX335 #42335 pX458 #48138 pX458M EZ-GuideXH pX459M EZ-GuideXH EZ-GuideXH pX461 #48140 pX552 #60958 pX601 #61591 pX602 #61593 LentiCRISPR V2 #52961 pLen
2015-9-11 · Plasmid Preparation. Vectors containing the human Cas9 and gRNA expression cassette pX260 and pX330 (Addgene) were used to create various constructs. Both vectors contain a humanized Cas9 coding sequence driven by a CAG promoter and a gRNA expression cassette driven by a human U6 promoter . The vectors were digested with BbsI and treated with
2019-9-2 · Plasmid. The cDNA encoding CLEC3B was achieved by PCR and cloned into the p3xFLAG-CMV-14 vector. The px260-CLEC3B was obtained as instruction. px260 plasmid was digested with Bbsl (Thermo scientific Fastdigest Bpi I FD1014) and a pair of oligos were annealed (Beyotime Biotechnology Shanghai China) which were cloned into the CRISPER array.
2019-9-2 · PX260 This plasmid separately encodes a human codon-optimized SpCas9 a tracrRNA and customizable crRNA. #42230 PX330 A human codon-optimized SpCas9 and chimeric guide RNA expression plasmid. #48138 PX458 GFP Cas9 from S. pyogenes
pX260 #42229 pX330 #42230 pX335 #42335 pX458 #48138 pX458M EZ-GuideXH pX459M EZ-GuideXH EZ-GuideXH pX461 #48140 pX552 #60958 pX601 #61591 pX602 #61593 LentiCRISPR V2 #52961 pLen
2016-3-4 · For gRNA expression screening specific reverse primer (pX260-crRNA-3′/R Table I.3 in S1 Experimental Procedures) was used in RT reaction followed by standard PCR using target A or B
a plasmid such as pX260 and pX335 which encodes both sgRNA and Cas9. The second delivery pattern is a mixture of Scheme 1 Key considerations in designing CRISPR/Cas9-carrying nanoparticles for therapeutic genome editing. Fig. 1 Delivery patterns of CRISPR/Cas9 components. The first delivery
2017-9-11 · For instance in the pX260 or pX330 system Cas9 protein and sgRNA were expressed form the same plasmid. The pX260 system also known as the pX334 system contains three cassettes including a CRISPR RNA array tracrRNA and S. pyogenes Cas9 (or the Cas9 D10A nickase).
BQ+ Medical not only produce components for manufacturers all over the world, but also provide private label manufacturing for leading brand distributors in different countries.
8,Cheye Road,Songjiang District,Shanghai 201611 China.
Tel: 86-021-57743953
Email: [email protected]
Copyright © BQ PLUS MEDICAL CO., LTDSitemap | Contact Us |
