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Gene expression knockdown through RNA interference(RNAi)is a widely used method for studying gene function. Small interfering RNA(siRNA)is commonly employed for transient knockdown in mammalian cell culture due to its quick and efficient transfection capabilities. However, siRNA has limitations in low transfection efficiency cell types and experiments requiring prolonged knockdown. An alternative method is short hairpin RNA(shRNA), a synthetic non-coding RNA utilizing endogenous microRNA machinery for functional RNAi. shRNA, forming a tight hairpin turn, silences gene expression via RNAi. Delivery can be achieved through plasmids, viral vectors, or bacterial vectors. Compared to siRNA, shRNA offers advantages in longevity, delivery options, and cost.
Creative Biogene provides a comprehensive collection of approximately 200, 000 pre-cloned shRNA constructs, targeting different regions of each gene sequence. Each shRNA is cloned into plasmids or lentiviral vectors, ensuring sequence verification for accurate target gene matching.
Key Features of Our shRNA Clones
shRNA(short hairpin RNA)has become a versatile molecular tool in biomedical research, enabling targeted gene silencing. Researchers use shRNA clones to study gene knockdown effects across diverse cell types and model organisms, providing insights into specific gene roles in cellular processes, disease development, and organismal physiology. This approach is extensively applied in functional genomics studies for systematic exploration of gene function on a genome-wide scale. Our products will drive your research on:
Case Study 1
Thioredoxin-interacting protein(TXNIP)is pivotal in oxidative stress, inflammation, apoptosis, and the development of diabetic retinopathy(DR). Researchers employed shRNA Clones to elucidate the role of TXNIP in high glucose-induced dysfunction of retinal pigment epithelium(RPE). High glucose(HG)significantly upregulated TXNIP expression in both ARPE-19 cells and primary human RPE(HRPE)cells, leading to mitochondrial membrane depolarization, fragmentation, and mitophagic flux to lysosomes. Confocal live-cell imaging revealed lysosomal enlargement and inactivation of cathepsin L under HG. TXNIP knockdown by shRNA prevented these effects, indicating its mediation of deleterious consequences of high glucose on RPE. Antioxidant N-acetylcysteine(NAC)and Amlexanox(Amlx)also mitigated mitophagic flux and lysosome enlargement. These findings highlight TXNIP's potential involvement in diabetic retinopathy development.
Figure 1. Researchers utilized shRNA Clones(shTXNIP 3+4)to efficiently
knock down TXNIP in ARPE-19 cells, employing stable cell selection with G417. Researchers performed TXNIP shRNA
clones transfection using pcDNA3. 1 plasmids(Cat#CBGT J0909-1, obtained from Creative Biogene, Shirley, USA). (Devi
TS,
et al. , 2022)
Case Study 2
Diabetic erectile dysfunction(ED)is on the rise, and the existing drugs are ineffective in treating it due to the severe angiopathy associated with diabetes. Researchers employed shRNA Clones to investigate the impact of insulin‐like growth factor‐binding protein 5(IGFBP5)knockdown on erectile function in diabetic mice. Diabetic conditions increased IGFBP5 expression in cavernous tissues and primary cavernous endothelial cells(MCECs). Knockdown of IGFBP5 enhanced MCECs angiogenic activity under high‐glucose conditions. In diabetic mice, shIGFBP5 treatment significantly improved erectile function, increased cell numbers, and positively influenced eNOS phosphorylation. IGFBP5 was identified as a mediator of AKT, ERK, and p38 signaling pathways. This research suggests local IGFBP5 inhibition as a potential strategy for treating diabetic erectile dysfunction and related vascular or neurological disorders.
Figure 2. Researchers utilized shRNA Clones, achieving IGFBP5
knockdown, restoring angiogenic activity in MCECsin vitroandex vivo. (Ock J, et al. ,
2023)
Q: What quality control measures are important for ensuring the effectiveness of shRNA constructs?
A: To ensure the effectiveness of shRNA constructs, rigorous quality control measures such as sequence verification are essential. Sequence verification ensures that the shRNA sequences accurately match the intended target genes, enhancing the specificity and reliability of gene knockdown experiments.
Q: How are shRNA constructs designed to minimize off-target effects while maximizing knockdown efficiency?
A: shRNA constructs are designed using advanced algorithms to optimize knockdown efficiency while minimizing off-target effects. This design process involves selecting target sequences within the gene of interest that are specific and conserved, reducing the likelihood of unintended gene silencing and enhancing the reliability of experimental results.
Q: In what experimental contexts can shRNA be applied for studying gene function?
A: shRNA can be applied in various experimental contexts to study gene function, including cellular biology, physiology, and disease development. Researchers can use shRNA to silence specific genes and investigate their roles in cellular processes, signaling pathways, and disease mechanisms.
Q: How can shRNA contribute to drug target screening and the development of gene-targeted therapies?
A: shRNA can be used for drug target screening by silencing specific genes associated with disease pathways. By assessing the effects of gene knockdown on cellular phenotypes, researchers can identify promising drug targets for further investigation and development of gene-targeted therapies. This approach expedites the drug discovery process and facilitates the development of precision medicine strategies.
| Cat.No. | Product Name | Price |
|---|---|---|
| SHG002873 | shRNA set against Mouse 1600012H06Rik(NM_026451.2) | Inquiry |
| SHG011599 | shRNA set against Mouse Apoa1(NM_009692.3) | Inquiry |
| SHG014959 | shRNA set against Mouse 4930579E17Rik(NM_178629.5) | Inquiry |
| SHG019689 | shRNA set against Human A1BG(NM_130786.3) | Inquiry |
| SHG020269 | shRNA set against Human A2M(NM_000014.4) | Inquiry |
| SHG020305 | shRNA set against Rat A2m(NM_012488.2) | Inquiry |
| SHG020773 | shRNA set against Rat A4galt(NM_022240.2) | Inquiry |
| SHG020791 | shRNA set against Human A4GALT(NM_017436.4) | Inquiry |
| SHG021589 | shRNA set against Mouse AA881470(NM_172724.3) | Inquiry |
| SHG021815 | shRNA set against Mouse Aacs(NM_030210.1) | Inquiry |
| SHG021833 | shRNA set against Human AADAC(NM_001086.2) | Inquiry |
| SHG021925 | shRNA set against Rat Aadat(NM_017193.1) | Inquiry |
| SHG022609 | shRNA set against Rat Abca1(NM_178095.2) | Inquiry |
| SHG022817 | shRNA set against Human ABCA2(NM_001606.4) | Inquiry |
| SHG023039 | shRNA set against Human ABCA3(NM_001089.2) | Inquiry |
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