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Expanding the range of editable targets in the wheat genome using the variants of the Cas12a and Cas9 nucleases

Expanding the range of editable targets in the wheat genome using the variants of the Cas12a and Cas9 nucleases

The event of CRISPR-based editors recognizing distinct protospacer adjoining motifs (PAMs), or having completely different spacer size/construction necessities broadens the vary of attainable genomic functions.
We evaluated the pure and engineered variants of Cas12a (FnCas12a and LbCas12a) and Cas9 for his or her capability to induce mutations in endogenous genes controlling necessary agronomic traits in wheat.
In contrast to FnCas12a, LbCas12a induced mutations within the wheat genome, though with a decrease fee than that reported for SpCas9. The eight-fold enchancment within the gene enhancing effectivity was achieved for LbCas12a through the use of the guides flanked by ribozymes and pushed by the RNA polymerase II promoter from switchgrass.
The effectivity of multiplexed genome enhancing (MGE) utilizing LbCas12a was principally just like that obtained utilizing the simplex RNA guides, and confirmed substantial enhance after subjecting transgenic crops to high-temperature therapy.
We efficiently utilized LbCas12a-MGE for producing heritable mutations in a gene controlling grain measurement and weight in wheat. We confirmed that the vary of editable loci within the wheat genome may very well be additional expanded through the use of the engineered variants of Cas12a (LbCas12a-RVR) and Cas9 (Cas9-NG and xCas9) that acknowledge the TATV and NG PAMs, respectively, with the Cas9-NG displaying larger enhancing effectivity on the targets with atypical PAMs in comparison with xCas9.
In conclusion, our examine studies a set of validated pure and engineered variants of Cas12a and Cas9 editors for focusing on loci within the wheat genome not amenable to modification utilizing the unique SpCas9 nuclease.

Genome engineering and illness modeling through programmable nucleases for insulin gene remedy; guarantees of CRISPR/Cas9 expertise

Focused genome enhancing is a regularly evolving expertise using programmable nucleases to particularly change, insert, or take away a genomic sequence of curiosity.
These superior molecular instruments embody meganucleases, zinc finger nucleases, transcription activator-like effector nucleases and RNA-guided engineered nucleases (RGENs), which create double-strand breaks at particular goal websites within the genome, and restore DNA both by homologous recombination within the presence of donor DNA or through the error-prone non-homologous end-joining mechanism.
A not too long ago found group of RGENs often known as CRISPR/Cas9 gene-editing methods allowed exact genome manipulation revealing a causal affiliation between illness genotype and phenotype, with out the necessity for the reengineering of the precise enzyme when focusing on completely different sequences. CRISPR/Cas9 has been efficiently employed as an ex vivo gene-editing device in embryonic stem cells and patient-derived stem cells to grasp pancreatic beta-cell improvement and performance.
RNA-guided nucleases additionally open the way in which for the technology of novel animal fashions for diabetes and permit testing the effectivity of assorted therapeutic approaches in diabetes, as summarized and exemplified on this manuscript.

Technology of Brachyury-mCherry knock-in reporter human pluripotent stem cell line (SNUe003-A-2) utilizing CRISPR/CAS9 nuclease

Brachyury is an embryonic nuclear transcription issue required for mesoderm formation and differentiation. Right here, we launched an mCherry reporter into the C-terminus of Brachyury within the human pluripotent stem cell line SNUhES3 utilizing the CRISPR/Cas9 nuclease strategy.
Profitable gene enhancing was verified by DNA sequencing. SNUhES3-Brachyury-mCherry cells expressed pluripotent stem cell markers, exhibited a standard karyotype, and will generate all three germ layers. This cell line expressed the purple fluorescence protein mCherry upon the induction of mesoderm differentiation.
This reporter cell line may very well be used to watch mesodermal inhabitants enrichment throughout mesodermal differentiation.

Software of CRISPR/Cas9 Nuclease in Amphioxus Genome Modifying

The cephalochordate amphioxus is a promising animal mannequin for finding out the origin of vertebrates resulting from its key phylogenetic place amongst chordates.
Though transcription activator-like effector nucleases (TALENs) have been adopted in amphioxus genome enhancing, its labor-intensive development of TALEN proteins limits its utilization in lots of laboratories.
Right here we reported an software of the CRISPR/Cas9 system, a extra amenable genome enhancing methodology, on this group of animals.
Our information confirmed that whereas co-injection of Cas9 mRNAs and sgRNAs into amphioxus unfertilized eggs prompted no detectable mutations at focused loci, injections of Cas9 mRNAs and sgRNAs on the two-cell stage, or of Cas9 protein and sgRNAs earlier than fertilization, can execute environment friendly disruptions of focused genes.
Among the many 9 examined sgRNAs (focusing on 5 genes) co-injected with Cas9 protein, seven launched mutations with effectivity starting from 18.4% to 90% and 4 prompted particular phenotypes within the injected embryos. We additionally demonstrated that monomerization of sgRNAs through thermal therapy or modifying the sgRNA construction may enhance mutation efficacies.
Our examine won’t solely promote software of genome enhancing methodology in amphioxus analysis, but additionally present invaluable experiences for different organisms by which the CRISPR/Cas9 system has not been efficiently utilized.

Exact Modifying of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice ( Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins

Abscisic acid (ABA) is concerned in regulating drought tolerance, and pyrabactin resistance-like (PYL) proteins are often known as ABA receptors. To elucidate the position of one of many ABA receptors in rice, OsPYL9 was mutagenized by means of CRISPR/Cas9 in rice.
Homozygous and heterozygous mutant crops missing any off-targets and T-DNA had been screened primarily based on site-specific sequencing and used for morpho-physiological, molecular, and proteomic evaluation.
Mutant strains seem to build up larger ABA, antioxidant actions, chlorophyll content material, leaf cuticular wax, and survival fee, whereas a decrease malondialdehyde stage, stomatal conductance, transpiration fee, and vascular bundles happen underneath stress circumstances.
Proteomic evaluation discovered a complete of 324 differentially expressed proteins (DEPs), out of which 184 and 140 had been up and downregulated, respectively. The OsPYL9 mutants confirmed a rise in grain yield underneath each drought and properly watered subject circumstances.
Many of the DEPs associated to circadian clock rhythm, drought response, and reactive oxygen species had been upregulated within the mutant crops.
Kyoto Encyclopedia of Genes and Genomes (KEGG) evaluation revealed that DEPs had been solely concerned in circadian rhythm and Gene Ontology (GO) evaluation confirmed that a lot of the DEPs had been concerned in response to abiotic stimulus, and abscisic acid-activated signaling pathways.
Protein GIGANTEA, Adagio-like, and Pseudo-response regulator proteins confirmed larger interplay in protein-protein interplay (PPI) community.
Thus, the general outcomes confirmed that CRISPR/Cas9-generated OsPYL9 mutants have potential to enhance each drought tolerance and the yield of rice.

Nuclease AWAY

A6257-100ML 100ML
EUR 22
Description: Biotechnology

Nuclease AWAY

A6257-500ML 500ML
EUR 88
Description: Biotechnology

Nuclease Mix 0.5mL

80650142 EACH
EUR 170.4

Water, Nuclease Free

42300012-1 1 L
EUR 60.1

Water, Nuclease Free

42300012-2 4 L
EUR 81.44

Water, Nuclease Free

42300012-3 125 mL
EUR 10.94

Water, Nuclease Free

42300012-4 250 mL
EUR 18.58

Water, Nuclease Free

42300012-5 500 mL
EUR 33.95

Water, Nuclease Free

42300012-8 8 L
EUR 173.72

Nuclease-free H2O

RK20102 25mL
EUR 21.5

Nuclease P1 Antibody

1-CSB-PA328459LA01PEU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is Unconjugated. Tested in the following application: ELISA

Nuclease P1 Antibody

20-abx300888
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Benzonase Nuclease, 99%

enz-1112 5000U
EUR 90
Description: Recombinant Serratia Marcescens Benzonase Nuclease, 99%

Benzonase Nuclease, 90%

enz-1150 5000U
EUR 70
Description: Recombinant Serratia Marcescens Benzonase Nuclease, 90%

Nuclease P1 Antibody (HRP)

20-abx300889
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Nuclease P1 Antibody (FITC)

20-abx300890
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Nuclease P1 Antibody (Biotin)

20-abx300891
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Bovine Albumin Nuclease Free

IBOALBNF100MG each
EUR 136
Description: Bovine Albumin Nuclease Free

Bovine Albumin Nuclease Free

IBOALBNF500MG each
EUR 347
Description: Bovine Albumin Nuclease Free

Universal Nuclease ELISA Kit

RK04660 96T
EUR 280

Nuclease Decontamination Solution

43094006-1 250 mL
EUR 43.46

Recombinant Human APEX nuclease-1

7-02476 2µg Ask for price

Recombinant Human APEX nuclease-1

7-02477 10µg Ask for price

Recombinant Human APEX nuclease-1

7-02478 1mg Ask for price

GENPower™ NLS-Cas9 Nuclease (MALS verified)

CA9-S5149 100ug
EUR 492.2
Description: GENPower™ NLS-Cas9 Nuclease (CA9-S5149) is expressed from E. coli cells for genome editing applications with CRISPR technology with N-terminus and C-terminus NLS for improved transport to the nucleus . It is ideal for in vitro and in vivo reactions. It contains AA Asp 2 - Asp 1368 (Accession # Q99ZW2-1).

Nuclease P1 Polyclonal Antibody

A64318
  • EUR 684.66
  • EUR 117.70
  • EUR 302.50
  • EUR 423.50
  • 100 µg
  • 20 ul
  • 50 ul
  • 100 ul

GENIUS™Nuclease DMF Filed

BEE-N3116 500KU
EUR 96.3
Description: GENIUS™Nuclease is a recombinant form of Serratia macescens extracellular endonuclease produced in Escherichia coli cells using a proprietary process at ACRObiosystems. GENIUS™Nuclease is a homodimer with monomer molecular masses about 30 kDa. Two disulfide bonds found in the nuclease are crucial to its activity and stability. The enzyme is a non-specific nuclease with high specific activity, which degrades both single- and double-stranded nucleic acids in any form ( single stranded, double stranded, linear, circular and supercoiled ). It hydrolyzes internal phosphodiester bonds present between the nucleotides to 5‘- phosphorylated oligonucleotides of 3-5 bases in length.

Penicillium citrinum Nuclease P1

1-CSB-EP328459PEU
  • EUR 733.20
  • EUR 370.80
  • EUR 2192.40
  • EUR 1126.80
  • EUR 1461.60
  • EUR 476.40
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
Description: Recombinant Penicillium citrinum Nuclease P1 expressed in E.coli

Penicillium citrinum Nuclease P1

1-CSB-EP328459PEUa0
  • EUR 733.20
  • EUR 370.80
  • EUR 2192.40
  • EUR 1126.80
  • EUR 1461.60
  • EUR 476.40
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
Description: Recombinant Penicillium citrinum Nuclease P1 expressed in E.coli

Penicillium citrinum Nuclease P1

1-CSB-YP328459PEU
  • EUR 814.80
  • EUR 402.00
  • EUR 2606.40
  • EUR 1261.20
  • EUR 1730.40
  • EUR 522.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
Description: Recombinant Penicillium citrinum Nuclease P1 expressed in Yeast

Cas9 Null Nuclease: EF1-hspCas9-DM-H1-gRNA NullNuclease vector

CAS805A-1 10 ug
EUR 724

anti-Apoptosis enhancing nuclease

YF-PA20583 50 ug
EUR 435.6
Description: Mouse polyclonal to Apoptosis enhancing nuclease

GENIUS™ Nuclease, premium grade

NUE-S5119 5000KU
EUR 1358.9
Description: GENIUS™ Nuclease, premium grade is a tag free recombinant form of Serratia marcescens extracellular endonuclease produced in Escherichia coli cells using a proprietary process at ACRObiosystems. GENIUS™ Nuclease, premium grade is a homodimer with monomer molecular masses about 30 kDa with an isoelectric point (pI) at pH 6.85. The enzyme is a non-specific nuclease with high specific activity, which degrades both single- and double-stranded nucleic acids in any form (single stranded, double stranded, linear, circular and supercoiled). It hydrolyzes internal phosphodiester bonds present between the nucleotides to 5’- phosphorylated oligonucleotides of 3-5 bases in length.

Rat APEX Nuclease 1 ELISA Kit

IRTAPEX1KT each
EUR 790
Description: Rat APEX Nuclease 1 ELISA Kit

Apoptosis enhancing nuclease antibody

70R-13466 100 ul
EUR 548.4
Description: Affinity purified Rabbit polyclonal Apoptosis enhancing nuclease antibody

Mouse APEX Nuclease 1 ELISA Kit

IMSAPEX1KT each
EUR 752
Description: Mouse APEX Nuclease 1 ELISA Kit

GMP GENIUS™Nuclease DMF Filed

GMP-NUES19 5000KU
EUR 2011.6
Description: GMP GENIUS™Nuclease DMF Filed is a tag free recombinant form of Serratia marcescens extracellular endonuclease produced in Escherichia coli cells using a proprietary process at ACRObiosystems. GMP GENIUSNuclease DMF Filed is a homodimer with monomer molecular masses about 30 kDa with an isoelectric point (pI) at pH 6.85. The enzyme is a non-specific nuclease with high specific activity, which degrades both single- and double-stranded nucleic acids in any form (single stranded, double stranded, linear, circular and supercoiled). It hydrolyzes internal phosphodiester bonds present between the nucleotides to 5’- phosphorylated oligonucleotides of 3-5 bases in length.

Human APEX Nuclease 1 ELISA Kit

IHUAPEX1KT each
EUR 733
Description: Human APEX Nuclease 1 ELISA Kit

Nuclease P1 Antibody, HRP conjugated

1-CSB-PA328459LB01PEU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is HRP conjugated. Tested in the following application: ELISA

NLS-Cas12a Nuclease (MALS verified)

CAA-L5149 100ug
EUR 107
Description: NLS-Cas12a Nuclease (CAA-L5149) is expressed from E. coli cells. It contains AA Ser 2 - His 1228 (Accession # QWN56480.1).

Apoptosis-enhancing nuclease antibody

22725-100ul 100ul
EUR 468

Nuclease P1 Antibody, FITC conjugated

1-CSB-PA328459LC01PEU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is FITC conjugated. Tested in the following application: ELISA

Fungus-Derived Nuclease S1 Purified

IFGS1NCL10KU each
EUR 100
Description: Fungus-Derived Nuclease S1 Purified

Fungus-Derived Nuclease S1 Purified

IFGS1NCL50KU each
EUR 247
Description: Fungus-Derived Nuclease S1 Purified

CAS9 CRISPR-Associated Protein-9 Nuclease S. Pyogenes Recombinant Protein

PROTQ99ZW2 Regular: 100ug
EUR 1832.4
Description: Recombinant Cas9-NLS produced in E.coli, comprises the entire Cas9 protein sequence (1368 amino acids) joined to a proprietary nuclear localization sequence (NLS) and a 6xHis tag at the C-terminus, having a total of 1414 amino acids. The protein has an apparent molecular weight of 163kDa.

Nuclease P1 Antibody, Biotin conjugated

1-CSB-PA328459LD01PEU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is Biotin conjugated. Tested in the following application: ELISA

Human Apoptosis-enhancing nuclease (AEN)

1-CSB-EP819883HU
  • EUR 456.00
  • EUR 256.80
  • EUR 1570.80
  • EUR 672.00
  • EUR 1047.60
  • EUR 314.40
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
Description: Recombinant Human Apoptosis-enhancing nuclease(AEN),partial expressed in E.coli

Rat APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Ra-48T 48T
EUR 658.8
Description: A sandwich quantitative ELISA assay kit for detection of Rat APEX Nuclease 1 (APEX1) in samples from tissue homogenates, cell lysates or other biological fluids.
Moreover, world proteome evaluation gives new potential biomarkers and understandings of the molecular mechanism of rice drought tolerance.

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