cas9 protein (New England Biolabs)

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New England Biolabs cas9 protein

IL-22 mediated VD-induced β-defensin expression in zebrafish intestine. (a) The deletion site by <t>CRISPR/Cas9</t> on the il22 gene exon (E)1 (exons are in blue boxes) was displayed. (b) The protein level of IL-22 in the intestine of WT and il22 -/- zebrafish was compared ( n = 6/group). The image is representative of 6 replicates. (c) Zebrafish at 3 mpf were i.p . injected with 10 7 CFU E. tarda or PBS, and the survival rate was recorded until 96 hours-post infection ( n = 10/group). (d) The gene expression of zfbd1 , zfbd2 and zfbd3 in zebrafish intestine was measured. (e) After WT and il22 mutant zebrafish at 2 mpf were fed with 0 or 800 IU/kg dietary VD 3 for 4 weeks, the transcript levels of zfbd1 , zfbd2 and zfbd3 in zebrafish intestine were evaluated ( n = 6–8/group). * p < 0.05, *** p < 0.001, ns: non-significance. See also Figures S2.

Cas9 Protein, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 565 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/zebrafish+codon+optimized+cas9+coding+sequence/EnGen+Spy+Cas9+NLS/pmc10012952-255-13-15
Average 99 stars, based on 565 article reviews

cas9 protein - by Bioz Stars, 2026-09

99/100 stars

Images

1) Product Images from "Vitamin D influences gut microbiota and acetate production in zebrafish ( Danio rerio ) to promote intestinal immunity against invading pathogens"

Article Title: Vitamin D influences gut microbiota and acetate production in zebrafish ( Danio rerio ) to promote intestinal immunity against invading pathogens

Journal: Gut Microbes

doi: 10.1080/19490976.2023.2187575

IL-22 mediated VD-induced β-defensin expression in zebrafish intestine. (a) The deletion site by CRISPR/Cas9 on the il22 gene exon (E)1 (exons are in blue boxes) was displayed. (b) The protein level of IL-22 in the intestine of WT and il22 -/- zebrafish was compared ( n = 6/group). The image is representative of 6 replicates. (c) Zebrafish at 3 mpf were i.p . injected with 10 7 CFU E. tarda or PBS, and the survival rate was recorded until 96 hours-post infection ( n = 10/group). (d) The gene expression of zfbd1 , zfbd2 and zfbd3 in zebrafish intestine was measured. (e) After WT and il22 mutant zebrafish at 2 mpf were fed with 0 or 800 IU/kg dietary VD 3 for 4 weeks, the transcript levels of zfbd1 , zfbd2 and zfbd3 in zebrafish intestine were evaluated ( n = 6–8/group). * p < 0.05, *** p < 0.001, ns: non-significance. See also Figures S2.

Figure Legend Snippet: IL-22 mediated VD-induced β-defensin expression in zebrafish intestine. (a) The deletion site by CRISPR/Cas9 on the il22 gene exon (E)1 (exons are in blue boxes) was displayed. (b) The protein level of IL-22 in the intestine of WT and il22 -/- zebrafish was compared ( n = 6/group). The image is representative of 6 replicates. (c) Zebrafish at 3 mpf were i.p . injected with 10 7 CFU E. tarda or PBS, and the survival rate was recorded until 96 hours-post infection ( n = 10/group). (d) The gene expression of zfbd1 , zfbd2 and zfbd3 in zebrafish intestine was measured. (e) After WT and il22 mutant zebrafish at 2 mpf were fed with 0 or 800 IU/kg dietary VD 3 for 4 weeks, the transcript levels of zfbd1 , zfbd2 and zfbd3 in zebrafish intestine were evaluated ( n = 6–8/group). * p < 0.05, *** p < 0.001, ns: non-significance. See also Figures S2.

Techniques Used: Expressing, CRISPR, Injection, Infection, Mutagenesis

Microinjection:

Article Title: Bacterial lipoate protein ligases rescue lipoylation and respiration deficiency in mammals
Article Snippet: .. The sgRNAs and Cas9-NLS protein (catalog number M0646T; NEB) were mixed 1:1 to a final mixture for microinjection. .. For LplA KI mice generation, linearized targeting vector (5-10ng/ul) will be added into Cas9/sgRNA mix for pronuclear microinjection, the linearized DNA was purified using DNA Clean & Concentrator ® -25 kit (Zymo research) and eluted in RNase-free water.

In Vitro:

Article Title: Genetic analyses suggest that larval and adult stages of Lucilia cuprina employ different sensory systems
Article Snippet: .. The gRNA cutting efficiency was tested in vitro using Cas9 protein [New England Biolabs (NEB), Ipswich, MA, USA, Cat. # M0646T]. ..

Purification:

Article Title: The white gene as a transgenesis marker for the cricket Gryllus bimaculatus
Article Snippet: Guide sequences sgRNA#63 (5′-GAGTTCCTGGAAGAATTAAA-3′) and sgRNA#66 (5′-GAAGGTCGTGTTGCATATCT-3′) were selected based on their predicted efficiency ( Moreno-Mateos et al. 2015 ). sgRNAs were synthesized using the EnGen sgRNA Synthesis Kit, S. pyogenes (New England BioLabs #E3322S) and purified using Monarch RNA Cleanup Kit (New England Biolabs #T2040L), following manufacturer's instructions. sgRNAs were stored as small aliquots at −70°C. .. EnGen Spy Cas9 NLS enzyme (New England BioLabs #M0646 M) (2.5 pmol/μl) and purified sgRNAs (2.5 pmol/μl each) were co-injected. ..

Concentration Assay:

Article Title: Distinct structural and functional connectivity of genetically segregated thalamoreticular subnetworks.
Article Snippet: Resource

CRISPR:

Article Title: Generation of Isogenic iPSC Lines for Studying the Effect of the p.N515del (c.1543_1545delAAC) Variant on MYBPC3 Function and Hypertrophic Cardiomyopathy Pathogenesis
Article Snippet: .. CRISPR/Cas9 RNP complexes were formed by incubation of 50 pmol of single-guide RNA (GenScript, Nanjing, China) and 20 pmol of Cas9_NLS (NEB, Ipswich, MA, USA) for 20 min at room temperature. .. The RNP complexes, together with 300 ng of single-stranded donor oligonucleotide (Biolegio, Nijmegen, The Netherlands), were delivered into 1 × 10 5 of patient-specific iPSCs (the ICGi029-A line, [ ]) by electroporation using Neon Transfection System 10 μL Kit (Thermo Fisher Scientific, Waltham, MA, USA) under the following conditions: 1100 V, 30 ms, one time.

Article Title: Generation of Isogenic iPSC Lines for Studying the Effect of the p.N515del (c.1543_1545delAAC) Variant on MYBPC3 Function and Hypertrophic Cardiomyopathy Pathogenesis.
Article Snippet: .. CRISPR/Cas9 RNP complexes were formed by incubation of 50 pmol of single-guide RNA (GenScript, Nanjing, China) and 20 pmol of Cas9_NLS (NEB, Ipswich, MA, USA) for 20 min at room temperature. .. The RNP complexes, together with 300 ng of singlestranded donor oligonucleotide (Biolegio, Nijmegen, The Netherlands), were delivered into 1 × 105 of patient-specific iPSCs (the ICGi029-A line, [29]) by electroporation using Neon Transfection System 10 μL Kit (Thermo Fisher Scientific, Waltham, MA, USA) under the following conditions: 1100 V, 30 ms, one time.

Incubation:

Article Title: Generation of Isogenic iPSC Lines for Studying the Effect of the p.N515del (c.1543_1545delAAC) Variant on MYBPC3 Function and Hypertrophic Cardiomyopathy Pathogenesis.
Article Snippet: .. CRISPR/Cas9 RNP complexes were formed by incubation of 50 pmol of single-guide RNA (GenScript, Nanjing, China) and 20 pmol of Cas9_NLS (NEB, Ipswich, MA, USA) for 20 min at room temperature. .. The RNP complexes, together with 300 ng of singlestranded donor oligonucleotide (Biolegio, Nijmegen, The Netherlands), were delivered into 1 × 105 of patient-specific iPSCs (the ICGi029-A line, [29]) by electroporation using Neon Transfection System 10 μL Kit (Thermo Fisher Scientific, Waltham, MA, USA) under the following conditions: 1100 V, 30 ms, one time.

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