Immunoprecipitation experiment

The interaction between DNA and protein bound by RNA: CHIRP

Immunoprecipitation experimentChromatin Isolation by RNA Purification (CHIRP) is a method for detecting the interactions between DNA and proteins bound to RNA in vivo, which can simultaneously analyze the interactions between IncRNA, circRNA, proteins, and DNA.

The significance of CHIRP experiment

IncRNA can interact with proteins, DNA, and RNA to achieve diverse functions. CHIRP can explore the functions and mechanisms of IncRNA interacting with DNA and proteins, making it a powerful tool for related scientific research. The study of IncRNA plays an important role in epigenetic regulation and other life activities, and CHIRP is a key area of research

IncRNA is an important technical tool for identifying RNA chromatin interactions across the entire genome and conducting qualitative, quantitative, and positional analyses.

CHIRP experimental principle

CHIRP (Chromatin Isolation by RNA Purification) is a nucleic acid probe that binds to biotin to pull down target RNA. The DNA chromosome fragments (DNA protein complexes) that interact with it are adsorbed and can be detected for RNA, DNA, and protein through qRT PCR, WB, sequencing analysis, and qualitative profiling.

Experimental process of CHIRP

① Probe design: Design an antisense transnucleotide probe:

② Cell culture;

③ Broken cells:

④ Hybridization of biotinylated DNA probes with RNA;

⑤ CHIRP;

⑥ Extraction of RNA, DNA, and nucleic acid binding proteins; .

⑦ Analysis and testing. ,

Results Display

Experimental completion cycle and delivery standards

1. Experimental period: 1-3 months

2. Delivery standards: CHIRP identification result report, analysis report, experimental report (reagents, consumables, experimental steps), raw data, etc

Information that needs to be confirmed with the customer for the experiment

1. Target gene and protein information.

2. Sample types and species

3. Is bioinformatics analysis necessary

References

[1]lyer S , Modali S D , Agarwal S K. The Long Non-Coding RNA MEG3 is an Epigenetic Determinant of OncogenicSignaling in Functional Pancreatic Neuroendocrine Tumor Cells[J]. Molecular and Cellular Biology, 2017:MCB.00278-17.

[2] Chu C , Chang H Y . Understanding RNA-Chromatin Interactions Using Chromatin Isolation by RNA Purification(ChIRP). [J]. Methods in Molecular Biology, 2016, 1480.

[3]ZhuJJ,FuHJ , Wu Y G , et al. Function of IncRNAs and approaches to IncRNA-protein interactions[J]. Science

China Life Sciences, 2013, 56(10):876-885.

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