Computational analysis and design of crrna for post transcriptional regulation of metastamir-10b and metastamir-126 by crispr-c2c2 technique.

Fatemeh Ebrahimi tarki,1,* Mahsa bourbour,2 Mahboobeh zarrabi,3

1. alzahra university,department of biotechnology, computational group
2. alzahra university,department of biotechnology, computational group
3. alzahra university, department of biotechnology,computational group

Abstract


Introduction

One of the important causes of death in cancer patients is metastasis. recently have been shown that specific group of micrornas, which are called metastamirs, has metastatic effects. have been identified that mir-126 is significantly correlated with the presence of early stage colorectal liver metastasis. in metastatic breast cancer mir-10b is overexpressed and the expression is induced by the transcription factor twist, which binds to the putative promoter of mir-10b. so reducing the level of these microns can prevent the metastatic events.

Methods

In 2017, the crispr-c2c2 has been represented as an effective technique for rna targeting. based on this ability, we can target microrna precursors. c2c2 protein has two rnase activites. but it needs a very precise gps to guide this protein for reaching its targets. c2c2 protein`s gps is called crispr rna (crrna). after crrna binding to c2c2, guide it to single strand rna. designing crrna for this technique is a very crucial step.without a specific crrna, crispr-c2c2 acts randomly. in this article we design crrna for targeting metastamir 10-b and metastamir-126 precursor by crispr rt, due to decreasing the transcript of them.

Results

Based on crrna structure and specificity, evaluation has been done on the number of transcript target sites and off-target numbers. the best one targets metastamir 10-b and metastamir-126 precursor with very high specificity

Conclusion

Our study showed that crispr-c2c2 can be used as a versatile tool in metastamir inhibition.it makes post transcriptional repression possible.by using designed crrna there is a high probability for making metastasis reduced or even stop.

Keywords

Crispr-c2c2, metastamir, metastasis inhibition, crrna, crispr-rt