The Impact of Computer Technology in Molecular Biology and ...
The Impact of Computer Technology in Molecular Biology and Genetics BY ABTAHI TISHAD TALK ABOUT GROWTH HORMONE Why this topic? I am a pre-medical student I am a research assistant at the UNC School of Medicine (Neuroscience)
PCR DNA Sequencing Cryostat-Sectioning I was mesmerized by the immense role of technology in data collection, storage, and analysis in the scientific world Overview: Computer Technology in Molecular Biology and Genetics Topics to Cover
Sequencing DNA Genomic Databases BLAST + Basic Algorithm Identifying Genes Significance of Genomic Analysis and Future Prospects Sequencing DNA
What is DNA sequencing? It is taking a sample of DNA and unraveling its nucleotide base sequences to be stored or analyzed Advances in technology has made DNA sequencing faster, cheaper, and more efficient than ever before. Cost of the Human Genome Project was $100 million is 2001 It was $1,245 in 2015 (DNA Sequencing, 2016)
How DNA Sequencing WorksSanger/Original Method 1. DNA segment broken into many fragments and extended by identical primers 2. Extension of DNA ends at selectively marked nucleotides, creating many different length strands 3. Smallest strand = 1st nucleotide, 4. 2nd smallest = 2nd nucleotide largest = last nucleotide 5. Final nucleotide of each strand is marked with a specific color coded dye to be recognized by device 6. Entire process is automated by the machine G
G T C A T A G C Next Generation Sequencing Better technological innovation has led to methods which can sequence DNA faster by many magnitudes Use of fluorescence imaging techniques to capture nucleotide sequences at a rate that can be 200 times faster than the original method!
Again 100% automated, but a lot faster Many techniques are essentially the Sanger method, but even more compartmentalized for efficiency Storing the Data: The GenBank The massive amounts of information gathered from DNA sequencing must be stored What is the GenBank? It is a very large database of DNA sequences (Klug, Cummings, Spencer Palladino, 2013)
Run by the National Center for Biotechnology Information(NCBI) It contains more than 100 billion base pair sequences of DNA, and it doubles in size every 14 to 18 months (Klug, Cummings, Spencer Palladino, 2013) GenBank (cont.) Are the DNA sequences just stored? No they are identified for the genes they code for and that sequence is assigned an accession number
This number is unique to a particular sequence Used to locate the sequence from the large database Connection to class: The use of algorithms Input: Accession Number Output: Target DNA sequence/gene of interest DNA vs. Genes DNA is the vector in which genetic material is stored.
Genes are specific DNA sequences which code for proteins Protein expression is what causes phenotypic traits (hair color, skin color, etc) Only 2% of human DNA consists of genes (Klug, Cummings, Spencer Palladino, 2013) How are the Genes Identified? Gene identification from DNA sequences is called annotation Annotation heavily relies on computer technology to ensure DNA
sequences are identified quickly, efficient, and correctly. BioInformatics What is BioInformatics? It is the use of computer hardware and software and mathematics to analyze large quantities of genetic information. Ex. The GenBank, BLAST A mesh of biology, computer science, and mathematics Basic Local Alignment Search Tool:
BLAST What is BLAST? It is a computer software which allows for comparison between DNA sequences in search of similarities. Why is this Useful? Unknown DNA sequences can be identified by comparing them to already known sequences in different species Ex. Using known DNA sequence in rats for Gene X to find a similar Gene Y in mice.
The sequences should have a high similarity Before Explaining BLAST: How DNA Works DNA consists of repeating patterns of nucleotides with 4 distinct nitrogenous bases: adenine, thymine, cytosine, and thymine The order and number of nitrogenous bases are what differentiates the DNA of different organisms Closely related organisms have very similar DNA sequences, and may even contain some of the same genes
How BLAST Works BLAST aligns two different DNA sequences and produces an identity value The identity value is a measure of similarity between the two sequences (Klug, Cummings, Spencer Palladino, 2013) Example: If DNA Sequence X in mice contains the insulin gene, and DNA sequence Y in rats is 99% similar to DNA Sequence X, then Y must contain the insulin gene, or a similar equivalent in rats
Thus, using BLAST we just identified the insulin gene in rats from known data about mice BLAST essentially uses the same mechanism as a primitive search engine The Basic Algorithm of BLAST BLAST works similar to most search engines Users have to input a DNA sequence, and/or the mRNA transcript of a specific gene within an organism The software contains an index of information (aka the GenBank database)
It reproduces results from the index to the user based on how similar they are to the initial search query The algorithm of the search can be changed by the user based on what they are looking for in particular BLAST Algorithm in CS-Speak 1. Removal of low complexity regions or repeats in query sequence 1. 2. 3.
4. Prevents false sense of high similarity Make a k letter word list of the query sequence 1. i.e. 3 letters 2. Speeds up the search process Scan database for matching words 1. Assign numerical values for results; i.e perfect match = 7, mismatch = -4 2.
Process is repeated for every word in query The matches are compiled together for statistical analysis based on matching score BLAST Algorithm (Cont.) 6. The matched sequences (HSP) which pass the statistical significance test are aligned together if they are continuous 7. These sequences are subsequently further analyzed and reported http://onlinelibrary.wiley.com/doi/10.1002/9780470015902.a0005253.pub2/full Demonstration Human actin Gene mRNA sequence Link 1: https://www.ncbi.nlm.nih.gov/gene
Link 2: https://blast.ncbi.nlm.nih.gov/Blast.cgi Potential Problems with the old BLAST Previous versions of BLAST did not account for Introns: non-coding sequences of DNA Location of promoters/enhancers/silencers
These are all needed to code a gene into a protein Two genes could contain similar nucleotide sequences, but code for different proteins Plus, BLAST requires you to have a template/already identified gene to compare the target gene to More Complex Organisms Require More Innovative Algorithms for Analysis There are simply more factors that need to be taken into account when study a complex organisms DNA such as a human, than when studying a less complex organism like a bacteria
How is this solved? Many annotation programs have adopted newer algorithms that incorporate many of the factors previously mentioned (Klug, Cummings, Spencer Palladino, 2013) Introns, promoters, enhancers/silencers, termination regionsetc BLAST now provides information on mRNA and protein sequences of various genes Lets Summarize: Computer Technology is Involved in Every Aspect of Genomic Analysis
DNA sequencing machines: Sequence DNA Online Databases: Store DNA sequences Computer Softwares: Analyze DNA sequences Without computers, much of todays knowledge in genetics would not be possible Significance of Genomic Analysis Identifying evolutionary relationship between different organisms
BLAST comparisons Basic gene to protein relationships Genomic analysis found that there are significantly more protein than there are genes that code for them Identifying the origin of genetic diseases and ways to cure/prevent them Availability of GMOs
Green Revolution in India saved the lives of millions of people through high yielding cereal crops Development of New Areas of Research Due to Tech Advances Nutrigenomics- The study of how diet affects the genes Allows scientists to hypothetically give subjects healthy dietary recommendations based on the analysis of their genes (Klug, Cummings, Spencer Palladino, 2013) Gene analysis is done through the help of technology to locate specific target sequences pertaining to health
Stone-Age Genomics The study of ancient DNA from primarily extinct animals (Klug, Cummings, Spencer Palladino, 2013) Allows for better understanding of the lives of organisms from the past, and similarities they have to organisms of today The Future: The Encyclopedia of DNA Elements Project (ENCODE) Successor of the Human Genome Project The goal is to identify all promoters, enhancers, silencers, and open reading frames within human genome (Klug, Cummings, Spencer Palladino, 2013) The Future: Personalized Genome Projects
The low cost of DNA sequencing due to newer technology has made it a possibility for individuals to get a read out of their own genome (Klug, Cummings, Spencer Palladino, 2013) Can lead to personalized medicine and better treatment of diseases Potential Concerns: Genetic Discrimination Technology has enabled us to learn more about our genome than ever before, but this can lead to potential concerns such as genetic discrimination. Current laws do not protect workers within small businesses, nor
anyone within the U.S. military (Genetic Discrimination, 2016) Hacking of sensitive genetic information With greater progress comes greater responsibility Works Cited DNA Sequencing. (n.d.). Retrieved March 30, 2017, from https:// www.khanacademy.org/science/biology/biotech-dna-technology/dna-se quencing-pcr-electrophoresis/a/dna-sequencing Genetic Discrimination. (2016, May 2). Retrieved March 30, 2017, from
https://www.genome.gov/10002077/ Klug, W., Cummings, M., Spencer, C., & Palladino, M. (2013). Chapters 18.3-18.5. In Essentials of Genetics (8th ed., pp. 362-372). Boston, MA: Pearson. Altschul, S. F. (2014). BLAST Algorithm. ELS. doi:10.1002/9780470015902.a0005253.pub2
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