miR-CLIP
MicroRNA Crosslinking and Immunoprecipitation
miR-CLIP was developed to identify mRNA targets of miRNA, using synthetic miRNAs (Imig et al., 2014). The capture probes in miR-CLIP are pre-miRNAs conjugated to psoralen and biotin that can be crosslinked with mRNA to isolate their targets from total RNA.First, a miR-CLIP capture probe needs to be designed and tested for functionality and strong binding affinity. Next, they are transfected into cells and UV-crosslinked at 254 nm and 365 nm, followed by immunoprecipitation of the crosslinked complex. The RNA is extracted and further purified by streptavidin pull-down. The isolated RNA targets can now be reverse-transcribed and processed into cDNA libraries for sequencing.
Advantages:
- Identifies targets of miRNAs genome-wide
- Capture probe miRNA sequences can be easily modified to investigate different miRNA species
Disadvantages:
- Not yet adopted widely by the scientific community
- Requires careful design of miRNA capture probes
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Sandhu G. K., Milevskiy M. J. G., Wilson W., Shewan A. M. and Brown M. A. Non-coding RNAs in Mammary Gland Development and Disease. Non-coding RNA and the Reproductive System. 2016;121-153
Yu B. and Shan G. Functions of long noncoding RNAs in the nucleus. Nucleus. 2016;7:155-166
Jeker L. T. and Marone R. Targeting microRNAs for immunomodulation. Curr Opin Pharmacol. 2015;23:25-31
Tycowski K. T., Guo Y. E., Lee N., et al. Viral noncoding RNAs: more surprises. Genes Dev. 2015;29:567-584
References:
Imig J., Brunschweiger A., Brummer A., et al. miR-CLIP capture of a miRNA targetome uncovers a lincRNA H19-miR-106a interaction. Nat Chem Biol. 2014;advance online publication:
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History: miR-CLIP
Revision by sbrumpton on 2017-06-21 07:50:24 - Show/Hide
MicroRNA Crosslinking and Immunoprecipitation
miR-CLIP was developed to identify mRNA targets of miRNA, using synthetic miRNAs (Imig et al., 2014). The capture probes in miR-CLIP are pre-miRNAs conjugated to psoralen and biotin that can be crosslinked with mRNA to isolate their targets from total RNA.First, a miR-CLIP capture probe needs to be designed and tested for functionality and strong binding affinity. Next, they are transfected into cells and UV-crosslinked at 254 nm and 365 nm, followed by immunoprecipitation of the crosslinked complex. The RNA is extracted and further purified by streptavidin pull-down. The isolated RNA targets can now be reverse-transcribed and processed into cDNA libraries for sequencing.
Advantages:- Identifies targets of miRNAs genome-wide
- Capture probe miRNA sequences can be easily modified to investigate different miRNA species
Disadvantages:- Not yet adopted widely by the scientific community
- Requires careful design of miRNA capture probes
Reagents:Illumina Library prep and Array Kit SelectorReviews:Sandhu G. K., Milevskiy M. J. G., Wilson W., Shewan A. M. and Brown M. A. Non-coding RNAs in Mammary Gland Development and Disease. Non-coding RNA and the Reproductive System. 2016;121-153Yu B. and Shan G. Functions of long noncoding RNAs in the nucleus. Nucleus. 2016;7:155-166Jeker L. T. and Marone R. Targeting microRNAs for immunomodulation. Curr Opin Pharmacol. 2015;23:25-31Tycowski K. T., Guo Y. E., Lee N., et al. Viral noncoding RNAs: more surprises. Genes Dev. 2015;29:567-584References:Imig J., Brunschweiger A., Brummer A., et al. miR-CLIP capture of a miRNA targetome uncovers a lincRNA H19-miR-106a interaction. Nat Chem Biol. 2014;advance online publication: