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

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.

Cas9 Nuclease

EN301-02 250 pmol
EUR 154

Cas9 Nuclease Lentiviral Vector

K002 10 ug
EUR 154

Cas9 Nuclease - 80µg - 1,600 ng/µl

3273-(HC) 1/EA
EUR 189

Cas9 Nuclease - 400µg - 1,600 ng/µl

3276-(HC) 1/EA
EUR 565

Nuclease P1 Antibody

1-CSB-PA328459LA01PEU
  • EUR 317.00
  • EUR 335.00
  • 100ug
  • 50ug
  • Form: Liquid
  • Buffer: Preservative: 0.03% Proclin 300
    Constituents: 50% Glycerol, 0.01M PBS, PH 7.4 >95%, Protein G purified
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 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Cas9 monoclonal antibody [Clone 7A9]

CAS9-AB-2 25 ug
EUR 204
  • Category: Cas9

A549 / Cas9 (Puro) Stable Cell Line

SC043-Cas9-Puro 2 x 106 cell/ml x 1ml
EUR 1058
Description: Cas9 expression stable cell line with Puromycin marker in A549 human lung cancer cells.

Hela / Cas9 (Bad) Stable Cell Line

SC045-Cas9-Bsd 2 x 106 cell/ml x 1ml
EUR 1058
Description: Cas9 expression stable cell line with Blasticidin resistance in Hela human cervical cancer cells.

A549 / Cas9 (GFP-Puro) Stable Cell Line

SC043-Cas9-GP 2 x 106 cell/ml x 1ml
EUR 1500
Description: Cas9 expression stable cell line with GFP-Puromycin fusion dual marker in A549 human lung cancer cells.

A549 / Cas9 (RFP-Puro) Stable Cell Line

SC043-Cas9-RP 2 x 106 cell/ml x 1ml
EUR 1500
Description: Cas9 expression stable cell line with RFP-Puromycin fusion dual marker in A549 human lung cancer cells.

Apoptosis-enhancing nuclease antibody

22725-100ul 100ul
EUR 390

Apoptosis enhancing nuclease antibody

70R-13466 100 ul
EUR 457
Description: Affinity purified Rabbit polyclonal Apoptosis enhancing nuclease antibody

Penicillium citrinum Nuclease P1

1-CSB-EP328459PEU
  • EUR 611.00
  • EUR 309.00
  • EUR 1827.00
  • EUR 939.00
  • EUR 1218.00
  • EUR 397.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 43.2 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Penicillium citrinum Nuclease P1 expressed in E.coli

Penicillium citrinum Nuclease P1

1-CSB-EP328459PEUa0
  • EUR 611.00
  • EUR 309.00
  • EUR 1827.00
  • EUR 939.00
  • EUR 1218.00
  • EUR 397.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 33.2 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Penicillium citrinum Nuclease P1 expressed in E.coli

Penicillium citrinum Nuclease P1

1-CSB-YP328459PEU
  • EUR 679.00
  • EUR 335.00
  • EUR 2172.00
  • EUR 1051.00
  • EUR 1442.00
  • EUR 435.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 31.2 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Penicillium citrinum Nuclease P1 expressed in Yeast

Nuclease P1 Antibody (HRP)

20-abx300889
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Nuclease P1 Antibody (FITC)

20-abx300890
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Nuclease P1 Antibody (Biotin)

20-abx300891
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Nuclease P1 Polyclonal Antibody

A64318 100 µg
EUR 570.55
Description: reagents widely cited

anti-Apoptosis enhancing nuclease

YF-PA20583 50 ug
EUR 363
Description: Mouse polyclonal to Apoptosis enhancing nuclease

Cas9 RT-PCR Primer Set (50ul at 5uM for each primer set)

CAS9-PR-1 50 reactions
EUR 153
  • Category: Cas9

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

Human Apoptosis-enhancing nuclease (AEN)

1-CSB-EP819883HU
  • EUR 380.00
  • EUR 214.00
  • EUR 1309.00
  • EUR 560.00
  • EUR 873.00
  • EUR 262.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 40.1 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Human Apoptosis-enhancing nuclease(AEN),partial expressed in E.coli

Nuclease P1 Antibody, HRP conjugated

1-CSB-PA328459LB01PEU
  • EUR 317.00
  • EUR 335.00
  • 100ug
  • 50ug
  • Form: Liquid
  • Buffer: Preservative: 0.03% Proclin 300
    Constituents: 50% Glycerol, 0.01M PBS, PH 7.4 >95%, Protein G purified
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is HRP conjugated. Tested in the following application: ELISA

Nuclease P1 Antibody, FITC conjugated

1-CSB-PA328459LC01PEU
  • EUR 317.00
  • EUR 335.00
  • 100ug
  • 50ug
  • Form: Liquid
  • Buffer: Preservative: 0.03% Proclin 300
    Constituents: 50% Glycerol, 0.01M PBS, PH 7.4 >95%, Protein G purified
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is FITC conjugated. Tested in the following application: ELISA

Nuclease P1 Antibody, Biotin conjugated

1-CSB-PA328459LD01PEU
  • EUR 317.00
  • EUR 335.00
  • 100ug
  • 50ug
  • Form: Liquid
  • Buffer: Preservative: 0.03% Proclin 300
    Constituents: 50% Glycerol, 0.01M PBS, PH 7.4 >95%, Protein G purified
Description: A polyclonal antibody against Nuclease P1. Recognizes Nuclease P1 from Penicillium citrinum. This antibody is Biotin conjugated. Tested in the following application: ELISA

Apoptosis-Enhancing Nuclease (AEN) Antibody

20-abx308819
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

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

CAS805A-1 10 ug
EUR 805
  • Category: Cas9

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

PROTQ99ZW2 Regular: 100ug
EUR 1527
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.

Fanconi-Associated Nuclease 1 (FAN1) Antibody

20-abx113962
  • EUR 732.00
  • EUR 398.00
  • 150 ul
  • 50 ul
  • Shipped within 5-10 working days.

Apoptosis-Enhancing Nuclease (AEN) Antibody (HRP)

20-abx308820
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Apoptosis-Enhancing Nuclease (AEN) Antibody (FITC)

20-abx308821
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Apoptosis-Enhancing Nuclease (AEN) Antibody (Biotin)

20-abx308822
  • EUR 411.00
  • EUR 1845.00
  • EUR 599.00
  • EUR 182.00
  • EUR 300.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug
  • Shipped within 5-10 working days.

Rabbit Reticulocyte Lysate System (Nuclease treated)

20-abx800017
  • EUR 9223.00
  • EUR 33096.00
  • EUR 5924.00
  • 100 ml
  • 500 ml
  • 50 ml
  • Shipped within 1 month.

Nuclease P1 Polyclonal Antibody, HRP Conjugated

A64319 100 µg
EUR 570.55
Description: Ask the seller for details

Nuclease P1 Polyclonal Antibody, FITC Conjugated

A64320 100 µg
EUR 570.55
Description: The best epigenetics products

Nuclease P1 Polyclonal Antibody, Biotin Conjugated

A64321 100 µg
EUR 570.55
Description: kits suitable for this type of research

GeneArt CRISPR Nuclease Vector- OFP- gRNA

PVT10418 2 ug
EUR 301

Human APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Hu-48T 48T
EUR 517
  • Should the Human APEX Nuclease 1 (APEX1) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Human APEX Nuclease 1 (APEX1) in samples from tissue homogenates, cell lysates or other biological fluids.

Human APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Hu-96T 96T
EUR 673
  • Should the Human APEX Nuclease 1 (APEX1) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Human APEX Nuclease 1 (APEX1) in samples from tissue homogenates, cell lysates or other biological fluids.

Mouse APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Mu-48T 48T
EUR 527
  • Should the Mouse APEX Nuclease 1 (APEX1) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse APEX Nuclease 1 (APEX1) in samples from tissue homogenates, cell lysates or other biological fluids.

Mouse APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Mu-96T 96T
EUR 688
  • Should the Mouse APEX Nuclease 1 (APEX1) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse APEX Nuclease 1 (APEX1) in samples from tissue homogenates, cell lysates or other biological fluids.

Rat APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Ra-48T 48T
EUR 549
  • Should the Rat APEX Nuclease 1 (APEX1) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
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.

Rat APEX Nuclease 1 (APEX1) ELISA Kit

DLR-APEX1-Ra-96T 96T
EUR 718
  • Should the Rat APEX Nuclease 1 (APEX1) ELISA Kit is proven to show malperformance, you will receive a refund or a free replacement.
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.

Bacillus subtilis Sporulation-specific extracellular nuclease (nucB)

1-CSB-YP337282BRJ
  • EUR 795.00
  • EUR 451.00
  • EUR 2288.00
  • EUR 1167.00
  • EUR 1558.00
  • EUR 551.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 12 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Bacillus subtilis Sporulation-specific extracellular nuclease(nucB) expressed in Yeast
Moreover, world proteome evaluation gives new potential biomarkers and understandings of the molecular mechanism of rice drought tolerance.

Leave a Reply

Your email address will not be published. Required fields are marked *