The RNA World GK-12 Presenation R.S. Michael Abdelsayed C.T. Mrs. Rojas NSF GK-12 NSF Grant DGE

Activities and Objectives Transcription – Students will make DNA into RNA Students should be able to understand that DNA is made into RNA Splicing – The RNA they make will splice itself Students should be able to understand that RNA is spliced to make a protein Ribozyme activity – Splicing will occur due to ribozyme! Students should be able to understand that in addition to proteins, RNA is an enzyme

California State Standards

Materials

Lab Prep

Question/Answers

Suggestions 2 Day Lab – Lecture first day (maybe demo gel), run lab second day Do lab after they have learned to make an agarose gel, or teach them during the lab (down time) Can tie lab in with enzymes or central dogma (transcription)

Central dogma DNA is made (transcribed) into RNA RNA is made (translated) into protein

DNA is made into RNA A protein enzyme, polymerase, transcribes (makes) DNA into RNA. You will transcribe DNA into RNA today!

RNA made into protein RNA is genetic material that codes for protein RNA must contain the correct code to make the correct protein

RNA Viruses!

RNA silencing RNA can turns genes off Normal plant Color blocked by RNA Color blocked by RNA

RNA Structure Non-linear RNA Linear mRNA tRNA

Create Your Own RNA

Ribozyme RNA that is an enzyme Enzymes speed up a reaction Ribozymes cleave (cut) RNA RNA Ribozyme

Splicing Splice – to join together at the ends Genes can be cut and pasted together (spliced!)

RNA Can Splice Itself Nobel Prize Thomas Cech RNA Ribozyme

Splicing Before splicing Exon 1 Intron Exon 2 After splicing Exon 1 Exon 2 Exon –codes for protein Intron – does not code for protein, splice it out!

Why Splice? Before splicing Not ready to be made into protein After splicing Ready to be made into protein Imagine someone gave you their phone number with ten extra numbers in the middle!

You will make RNA! The RNA YOU make will splice itself The RNA you make will cut itself and put itself together RNA Ribozyme

Procedure 9uL DNA Add 1uL Polymerase transcription enzyme Transcribe (make RNA) Incubate at 37 degrees DNA + Polymerase = RNA Ribozyme

Where does the ribozyme come from? Its will come from the RNA you make. The ribozyme is in the intron and cuts itself out ! RNA Ribozyme

Results

What it will look like DNARNA

Possible Adjustments Present as a ribozyme (enzyme) lab only (no transcription or splicing mentioned) “Today we will cut RNA” Protocol remains the same Incubate RNA, then observe on gel that RNA cuts itself

Enzyme

RNA is an enzyme, Ribozyme RNA that is an enzyme Enzymes speed up a reaction Ribozymes cleave (cut) RNA RNA Ribozyme

Enzymes – Temperature, ions, pH All enzymes have a temperature and a pH at which they work the fastest. Ions can change how well an enzyme works Enzymes in human cells work best at 37 o C (or 98.6 o F). – Maybe why our bodies are always 98.6??

Procedure 9uL DNA Add 1uL Polymerase transcription enzyme Transcribe (make RNA) Incubate at 37 degrees DNA + Polymerase = RNA Ribozyme

Procedure Incubate at 37 degrees Optimal Temperature RNA Ribozyme Can change temperature (37°C works best) Can add MgCl2 (May work faster)

Results Cut RNA Not cut RNA

Thanks GK-12 Mrs. Rojas Dr. Luptak GK-12 Directors Dr. Mota-Bravo Dr. de la Cruz Dr. Mulligan NSF GK-12 NSF Grant DGE