Advanced Techniques in Biology & Medicine

Advanced Techniques in Biology & Medicine
Open Access

ISSN: 2379-1764

Conference Proceeding - (2021)Volume 9, Issue 8

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Mutation Detection via CRISPRCas9 Micro-fluid Chip

Umair Masood*
 
*Correspondence: Umair Masood, Department of Biology, COMSATS University Islamabad, Pakistan, Email:

Author info »

Abstract

In biology mutation is a change in the nucleotide sequences of the DNA of an organism mainly there are three types of mutation: point mutation, deletion and insertions. Once the mutation has been defined allele specific oligonucleotide hybridization, amplification, heteroduplex formation method referred to as a diagnostic method some advance technique like CRISPR cas9 system is using for selected mutagenesis. Using microfluid and CRISPR cas9 system we can detect a mutation. Let's say you have a DNA sample with fluorescent labeled from patient and you want to make sure that gene you are interested is in healthy gene. We can design a CRISPR to scan through DNA or find specific gene or mutation. The CRISPR scan the DNA if the CRISPR does not find targeted gene it does not bind to it its means that no fluorescence color appears under UV-light but its scan and find its target and this binding create a fluorescence signal its means that mutation can be occur in gene

Microfluid Crispr Chip Parts

CRISPR micro fluid contains three main channels:

1. Sample loader

2. Mixture

3. Crispr block

Sample Loader

Sample loader contain small hole in order to load your DNA sample

Mixture

The mixture contains a zig zag design in order to move a sample from loader to Crispr Block

Crispr Block

Crispr block contain a Cas9 protein and guide RNA.

Working Mechanism

1. Load a DNA sample

2. Moving a sample from loader to Crispr Block

3. Water bath or incubate 37°C for 20 min because of cas9 protein

4. Observe under UV light (Figures 1 & 2).

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Figure 1. Working principal of CRISPRCas9 micro-fluid chip.

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Figure 2. Microfluid crispr chip parts.

Result and Observation

As we know fluorescence are absorbed the UV-light and emitted color visible that we can see. When a DNA is completely bind with cas9 protein which is going to tested and emitting a high color fluorescence light its mean that there is mutation either single site or multiple site mutation [1-5].

References

  1. Mulepati S, Héroux A, Bailey S. Crystal structure of a CRISPR RNA-guided surveillance complex bound to a ssDNA target. Science. 2014;345(6203):1479-1484.
  2. Barrangou R. The roles of CRISPR–Cas systems in adaptive immunity and beyond Curr Opin Immunol. 2015;1(32):36-41.
  3. Cotton RG. Current methods of mutation detection. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis. 1993; 285(1):125-144.
  4. Chen J, Miller BF, Furano AV. Repair of naturally occurring mismatches can induce mutations in flanking DNA. E life. 2014;3:123-129.
  5. Amann R, Ludwig W. Ribosomal RNA-targeted nucleic acid probes for studies in microbial ecology. FEMS microbiology reviews. 2000;24(5):555-565.

Author Info

Umair Masood*
 
Department of Biology, COMSATS University Islamabad, Islamabad, Pakistan
 

Citation: Masood U (2021) Mutation Detection via CRISPRCas9 Micro-fluid Chip. Adv Tech Biol Med. 9:317

Received: 26-Jun-2021 Accepted: 13-Aug-2021 Published: 25-Aug-2021 , DOI: 10.35248/2379-1764.21.9.317

Copyright: © 2021 Masood U. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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