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Where da protein at? | CRISPR/Cas9 gene editing and subcellular protein localization Topic Teaching molecular biology techniques has become common practice in introductory biology laboratories at both the high school and college level. Introductory biology laboratories often integrate cookbook experiments in molecular technique, but fail to apply these methods in a realistic context. Performing disconnected experiments does not engage students in the process of science (Stiller 2016). Holy Names University students have minimal access to research laboratories and environments. Bringing modern technology and a research context to students would increase engagement and prepare them for opportunities and future careers. Knowledge of protein localization within the cellular environment is critical for understanding the function of the protein and its integration with the complex network of cellular activity. In collaboration with Barbara Panning’s Laboratory at UCSF, we are taking advantage of a stable transgenic line, a split GFP system (Hu 2011) and CRISPR/Cas9 technology to visualize protein localization (Ran 2013).The laboratory experiment proposed here is a semester-long project that mimics the experiences of research laboratory environments. CRISPR/Cas9 technology takes advantage of a protein found in bacteria that makes double-strand breaks at sequence directed locations in the genome (Figure 1). The location of the double strand break is designated by the sequence located in a portion of the protein called the guide RNA. After the double strand break, homology directed repair can replace existing genome sequence with target sequence. CRISPR/Cas9 genome editing is a fast, efficient, and inexpensive way to alter the genome.

Figure 1. DSB repair promotes gene editing. DSBs induced by Cas9 (yellow) can be repaired in one of two ways. In the error-prone NHEJ pathway, the ends of a DSB are processed by endogenous DNA repair machinery and rejoined, which can result in random indel mutations at the site of junction. Indel mutations occurring within the coding region of a gene can result in frameshifts and the creation of a premature stop codon, resulting in gene knockout. Alternatively, a repair template in the form of a plasmid or ssODN can be supplied to leverage the HDR pathway, which allows high fidelity and precise editing. Single-stranded nicks to the DNA can also induce HDR. (figure and description from Ran 2018)

In order to perform these experiments, one would need to make a plasmid with the appropriate guide RNA sequences (this plasmid would also promote Cas9 expression and carry puromycin resistance). A second plasmid would be made to carry gene-specific exon and 3’UTR sequences plus a portion of a fluorescent tag, a “split GFP” fused to the C-terminus, for homology directed repair. The split GFP separates GFP into two separate proteins that associate with each other in vivo: GFP1-10 includes barrels 1-10 of the beta-barrel and GFP11 includes the 11th barrel (Kamiyama 2016). The stable transgenic mouse embryonic stem cell line expresses the GFP1-10 under control of a strong, constitutive promoter (TIGRE). Because the DNA sequence for GFP11 is only about 54 nucleotides, we can design oligos to easily subclone short homology arms and incorporate that 11th barrel into the 3’ end of a gene (5’-end targeting is possible but comes with additional considerations for gene structure). Expression of both pieces of the GFP protein leads to assembly of a complete GFP protein fused to the protein produced by the gene of interest (Figure 2). This GFP can be visualized through fluorescent microscopy and documented.

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Figure 2. Schematic diagram for fluorescent protein (FP) tag. FP11 is fused to the protein of interest and FP1-10 is expressed under control of a strong, constitutive promoter (Figure from Kamiyama 2016)

Purpose ● The scientific purpose of the laboratory experiment is to identify the subcellular localization of

proteins and better understand structure, function, and interactions with other proteins. ● The educational purposes of the laboratory are to

a. engage students by integrating and applying bioinformatic, molecular biology, and genetic techniques into one continuous research project and

b. Expose students to an experience similar to that of research laboratories ● Specifically at the end of the laboratory, students will be able to

a. Search bioinformatic databases for gene sequences b. Identify the function of gene components including the promoter, 5’-UTR, intron, exon, 3-

UTR, start codon, stop codon, and signal peptide sequences c. Design guide RNA constructs and PCR primers to allow CRISPR/Cas9 system to

incorporate GFP11 into genes of interest via homology-directed repair d. Perform molecular biology techniques (PCR, agarose gel electrophoresis, ligation,

transformation, plasmid purification) e. Culture and transfect mammalian cells using aseptic techniques f. Screen for GFP-positive cells using fluorescent microscopy g. Discuss ethical issues around CRISPR/Cas9 technology

Description The full laboratory sequence described here, is intended to be utilized in a 2 unit, upper division laboratory that meets for 6 hours per week at Holy Names University (Table 1). The funds from the Roberta Williams Laboratory Teaching Initiative Grant money would be utilized to develop positive control plasmids for 8 proteins with different cellular localization patterns (diffuse nuclear, nuclear and associated with DNA, cytoskeleton: actin, tubulin, lamin, diffuse cytoplasmic, mitochondrial, and nuclear membrane) and allow for troubleshooting the protocols on the Holy Names University campus. Portions of the laboratory will initially be tested through courses or mentored research experiences. The full sequence described in Table 1 could be shortened or expanded for different course timing and goals. For example, the instructor could perform some of the techniques and/or build and maintain a library of plasmids for students to investigate instead of having students build a plasmid from scratch. Table 1. Full semester laboratory sequence

Week Activity

1 Overview of laboratory sequence, CRISPR, & visualization of GFP-positive control samples (instructor would set up samples ahead of time).

2. Transformation of Positive control plasmids. Identify a gene/protein of interest and begin bioinformatic and background exploration

3 Liquid E.coli cultures, Cell-culture techniques

4 Mini-Preps, DNA Quantitation, Restriction digests

5. Agarose Gels, Cell-culture techniques

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6. Oligonucleotide design and order

7. PCR, cell-culture techniques

8. Agarose Gel and Gibson Ligation, cell-culture techniques

9. Transformation, Cell-culture techniques

10. Liquid Cultures, Cell-culture techniques

11. Mini-Preps, DNA Quantitation, Restriction digests

12. Agarose Gels, prepare for experiment

13. Cell culture techniques & Transfection

13b. Depending on course timing and student schedules, instructor will apply puromycin selection agent

14. Visualize and image cells

15. Optional: subculture cells

Example of student work assessment criteria In order to assess students in this laboratory activity, we would utilize the following activities:

1. To test students ability to utilize bioinformatics resources and the fundamentals of subcloning, there will be a culminating assignment where students would outline the process for constructing plasmids needed for an additional novel protein of interest. Students would design guide RNA’s and primers for PCR amplification of the HDR domains “in silico”.

2. To test students ability to perform aseptic techniques in the biosafety hood, we would observe them passaging cells and assess aseptic cell culture technique via a checklist.

3. To tests students ability to communicate the experimental context, methods, results, and implications, we would assign a culminating laboratory report that mimics journal article format (established rubric available upon request) and/or a poster presentation (established rubric available upon request).

4. We also plan to set up a website where students can view previous semester’s work and build an ongoing project resource.

An explanation of how the project will be sustained after the termination of ABLE funding In order to sustain this project after termination of ABLE funding, we will utilize the current budget for upper-division laboratory course supplies at HNU ($1,000). After the initial set-up funds, the estimated cost of running a laboratory with 20 students working as 10 pairs is less than $1000 (estimated at $725). Many purchased supplies will lead to “leftovers” and partially used materials that can carryover from semester to semester as applicable. An explanation of how the proposal clearly reflects current safety standards and animal use protocols

● All students will be trained in general laboratory safety. ● Mouse embryonic stem cells are classified as Biosafety Level 1. The laboratory facilities and

biosafety hoods where the work will take place are certified for Biosafety Level 2 work. ● Students will be trained in proper handling of materials and personal protective equipment for

BSL1 and BSL2 cell culture work. ● Personal protection equipment will be utilized including a lab coat, gloves (latex or nitrile as

appropriate), and safety glasses. ● Biohazard waste will be autoclaved and disposed of according to institutional policies. ● No vertebrate animal work will be involved with this protocol.

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Budget The budget items requested from this grant are divided into three sections:

● Table 2 outlines laboratory supplies that are needed to develop the 8 sets of positive control constructs. These items are also what would need to be purchased to run one section of the laboratory. The total for these items is approximately $725 (oligo length and price can vary).

● Table 3 outlines laboratory supplies that would be needed for a one-time purchase or purchased infrequently (once every 5-6 years).

● Table 4 outlines laboratory supplies that are readily available at HNU or through collaboration with the Panning Lab at UCSF.

Table 2. Purchases for laboratory development (and items that would need to be purchased for each lab section in the future)

Item Price Justification

Oligonucleotides for designing guide crRNA’s $100 $5 per guide RNA (about 20-25bp), 4

oligo’s per gene, 8 genes

Oligo’s for adding 11th barrel into HDR via PCR & Gibson Ligation $240

$5 per primer, 4-6 primers per gene, 8 genes

Hi Fidelity Gibson Ligation Materials $159

Most efficient way to subclone for HDR plasmid, a "one-step" procedure to

increase student success

DMEM Culture Media $150 Media for maintaining mouse embryonic

stem cells

Qiagen miniprep (50 reactions) $100

Only positive clones will be isolated using the Qiagen miniprep kit. Generally

bacterial clones will be screened using a "quick-mini-prep" protocol. Note: for a course, there would be carryover to the

next semester of this reagent.

Transfection Reagent $150

For transfecting cells. Note: for a course, there would be carryover to the next

semester of this reagent.

PCR Master Mix $79

Utilized for amplification of HDR sequence. Note: for a course, there

would be carryover to the next semester of this reagent.

Table 3. One-time purchases for course development

Item Price Justification

Fluorescent microscope with filter set for visualization GFP. $2,500

In order to perform the culminating experiment, we would need a fluorescent microscope. Currently HNU does not have a working fluorescent microscope.

RNAse A $50 For making "quick-mini prep" solutions to

screen bacterial colonies

Restriction Endonucleases $200

Many endonucleases are already available at HNU, we anticipate the need to purchase 3 additional enzymes due to the available sequences in plasmids for

subcloning.

Fetal Bovine Serum $150 Serum for maintaining mouse embryonic

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stem cells

Puromycin $250 Antibiotic utilized for selecting against

cells that were not transfected Table 4. Consumable materials from HNU and UCSF

Item Justification Source

Competent Cells (lab made) For transformations HNU

Bacteria/agar plates Media, Agar, antibiotic, Petri dishes HNU

Agarose gel electrophoresis material

Checking PCR and restriction digests HNU

Pen-Strep Antibiotic for maintaining growth of cells

Prevents contamination HNU

Tissue culture plates (i.e. 24 well, 6-well plates)

Provides a surface for cells to grow HNU

Fluorescent Microscope Maintenance

To keep microscope working/troubleshooting set-up

HNU

Plasmids For making guide RNA’s and homology directed repair constructs

Gift from Panning Lab

Leukemia Inhibitory Factor Needed for growth of embryonic stem cells

Gift from Panning Lab

Mouse Embryonic Stem Cells with stable incorporation of the 1-10 GFP barrel at the TIGRE locus

Cell-line that provides part of the GFP protein

Gift from Panning Lab

References Hu Y, Janitz M. High-throughput subcellular protein localization using transfected-cell arrays. Subcellular protein localization using cell arrays. Methods Mol Biol. 2011;706:53-72. Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert LA, Ishikawa H, Leonetti MD, Marshall WF, Weissman JS, Huang B. Versatile protein tagging in cells with split fluorescent protein. Nat Commun. 2016 Mar 18;7:11046. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Genome engineering using the CRISPR-Cas9 system. Nat Protoc. 2013 Nov;8(11):2281-2308. Stiller JW., Coggins TC. Teaching Molecular Biological Techniques in a Research Content. American Biology Teacher. Jan 2006; 68:36-42.

Department of Biochemistry &

Biophysics Barbara Panning, Professor Genentech Hall, S372B 600 16th Street San Francisco, CA 94143-2200 tel: 415/5 514-0745 fax: (415) 514-4080

University of California San Francisco

UCSF Biochemistry & Biophysics

April 29, 2018 Dear Association of Biology Laboratory Educators, I am Dr. Barbara Panning, Professor at University of California, San Francisco (UCSF). I support Dr. Chantilly Apollon’s application to the Roberta Williams Laboratory Teaching Initiative Grant. I have known Dr. Apollon since she was a graduate student at UCSF from 2002-2007. She recently reached out to me and invited me to Holy Names University (HNU) to speak to students. During the course of my visit, I learned about the types of equipment and resources available in the laboratory facilities. When we started talking about the laboratory teaching experiences Professor Apollon incorporates into the curriculum, I realized that my research laboratory could help bring more of the inquiry and research-based experiences to HNU students. I suggested she use a mouse embryonic stem cell line that we have developed in my laboratory that has part of GFP expressed constitutively as well as CRISPR/Cas9 technology. To aid the development of this laboratory activity, Dr. Apollon is welcome to work in my laboratory for a few weeks. In addition, my laboratory is happy to assist with reagents, such as the mouse embryonic stem cell lines and LIF, a protein required for stem cell growth. I know Dr. Apollon is striving to bring technology and research experiences to the HNU campus. She is deeply committed to her students and to developing a curriculum that will position them for jobs in the biotech sector. I am enthusiastic about supporting her in this endeavor. Sincerely,

Barbara Panning

Apollon

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CHANTILLY A. APOLLON*, PHD *née Munson 3500 Mountain Blvd. Oakland, CA 94619 e-mail: Apollon@hnu.edu office phone: 510-436-1413

EDUCATION

Ph.D. University of California, San Francisco September 2007 Developmental Biology Project: Analysis of endoderm organogenesis in zebrafish. Advisor: Didier Stainier, PhD B.S. University of Arizona - Tucson, AZ Spring 2002

Bachelor of Science, Biochemistry with Honors Bachelor of Science, Nutritional Sciences Summa Cum Laude

Minor in Spanish

TEACHING EXPERIENCE

Assistant Professor Holy Names University – Oakland, CA Math and Science Division, Biology Department Fall 2014 - present Special Topics in Biology: Cell Culture Techniques Spring 2018

• Course focusing on cell culture techniques including passaging cells, transfection, plasmid isolation, Dual Luciferase Reporter, and Western Blotting • Guest speakers from UCSF, SFSU presented on career pathways and research • Field trips to UCSF, SFSU, and Genentech Advanced Human Physiology (BIOL 115) Fall 2015 and Fall 2016

Exercise Physiology Laboratory (BIOL 115LW) • Laboratories focus on assessment of various components of fitness • Students developed unique projects for data collection and analysis • Incorporated electronic graphing and statistical analysis into weekly laboratory

exercises • Improved students’ scientific communication skills through a laboratory report

assessment intervention. Genetics (BIOL 160) Fall 2017

• Utilizing a genetics concept inventory to guide key topics and assess student learning • Incorporating laboratory experiences into a “lecture” course including Drosophila

mating and PCR

Principles of Biological Sciences (BIOL 1A and 1B) Spring 2016, 2017, 2018 • Utilizing hands-on activities during lecture to facilitate deeper learning of the

relationship between structure and function

Biochemistry: Physiological Chemistry (BIOL 185) Spring 2015, Spring 2017 Biochemistry: Physiological Chemistry Laboratory (BIOL 185L)

• Taught upper division biochemistry lecture and accompanying laboratory course focusing on biochemistry fundamentals and human metabolism

• Incorporated BIORAD Life Science Education Activities and developed novel extensions of these activities to better meet upper division laboratory objectives.

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• Incorporating novel research activities linked to zebrafish lateral line development and Wnt proteins during Spring 2017

• Taught students how to prepare video presentations on metabolic enzymes linked with common genetic disorders

Nutrition (NUTR 001) Spring 2015, 2016 & 2018, Fall 2015 • Teaching fundamentals of human nutrition and metabolism and including discussing of

current topics and controversies in the field of nutrition • Utilize student response system to collect formative assessment data • Course has been converted to a hybrid course (meeting face-to-face one day a week

and complete activities remotely one day a week) since Fall 2015 Human Physiology Lecture and Lab (BIOL 11) Fall 2014, Spring 2016

Human Physiology (BIOL 11) Fall 2015, Spring 2016 • Lecture and laboratory for students pursuing their Bachelor of Science in Nursing

degree • Utilize concept mapping and weekly online quizzes to promote long-term retention

and facilitate student comprehension of homeostasis • Incorporate electronic graphing and statistical analysis into weekly laboratory

exercises Tenured Instructor City College of San Francisco – San Francisco, CA 2007-2014 Biology Department Tenured since Fall 2011

Human Biology (BIO9) Fall 2007 – Spring 2014 ! Taught 2-3 sections of a combined lecture and laboratory course for non-majors each

semester ! Updated and improved laboratory exercises focusing on inquiry-based approaches ! Course coordinator for more than 10 sections each semester ! Incorporated technology such as iClickers and Moodle-based activities ! Developed hybrid version of course (students meet both online and face-to-face) ! Receive excellent student evaluations

General Biology Laboratory (BIO100A and BIO100B) Spring 2008 - present • Taught 1 section of a biology laboratory course for majors each semester • Updated several laboratory exercises in collaboration with colleagues

Adjunct Lecturer Skyline College - San Bruno, CA 2005-2007

Division of Science, Math and Technology

Human Biology (BIO 130) Fall 2006 & Spring 2007 ! Taught lecture course to about 40 non-majors for two semesters ! Was solely responsible for course content & instruction Introduction to Physiology (BIO 260) Spring & Fall 2005 ! Taught lecture and laboratory course to about 35 students each semester ! Was solely responsible for course content & instruction ! Evaluated highly by students who remarked upon my ability to effectively explain

difficult concepts

RESEARCH & MENTORSHIP EXPERIENCE

Visiting Professor San Francisco State University 01/2015 – present

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Laboratory of Laura Burrus, Ph.D. Project: Analysis of Wnt signaling and cholesterol metabolism in zebrafish lateral line

• Testing how Wnt signaling influences the developing zebrafish lateral line • Collaborative Animal Protocol for utilizing zebrafish at SFSU • Active participant in laboratory meetings, journal clubs, and data analysis

Directed Research Leader NIH Bridge to Baccalaureate Program SFSU Summers 2009 - present

• Lead 20 undergraduate community college students in 4 weeks of zebrafish research • Program targeted to support under represented minorities in the sciences in their pursuit of advanced science

degrees • Mentored students in individual projects examining zebrafish genetics and development • Data collected from external evaluator showed positive impact of program on students’ interest in pursing

advanced degrees and student enjoyment of having myself as a mentor • Program successfully launched many students to transfer and earn Bachelor’s Degrees, Master’s Degrees, and

Ph.D’s Graduate Student University of California, San Francisco

Tetrad Program 06/2003 – 08/2007 Advisor: Didier Stainier, Ph.D.

Project: Analysis of pard6gb in zebrafish organogenesis. ! Conducted independent developmental biology research. ! Applied molecular biology and live-imaging techniques to answer biological questions ! Collaborated with lab members and students from other laboratories in daily work. ! Trained undergraduate and visiting graduate students in laboratory techniques ! Trained graduate students and postdoctoral fellows in how to utilize the computer network and computer

programs. PROFESSIONAL ACTIVITIES

Coordinator, Kinesiology Program at Holy Names University Spring 2018 - present Peer Reviewer American Biology Teacher Spring 2017 - present Course Source Fall 2016 - present NSTA Journal of College Science Teaching Spring 2016 – present Expanding Your Horizons (EYH) Planning Committee Fall 2009 - Fall 2016

• Recruited presenters for hands-on workshops encouraging science careers for middle school girls Quality Matters Learning Community (Holy Names University) Fall 2014

• Worked with the Center for Teaching and Learning on applying the Quality Matters rubric to online courses

• Completed Quality Matters Certificate American Society for Microbiology – Biology Scholars Research Fellow 2013-2014

• Attended three day intensive workshop focusing on developing and implementing research questions in the college classroom

CCB-FEST (Community College Biology Faculty Enhancement through Scientific Teaching) 2009 - 2015

• Collaboration between San Francisco State University and Bay Area Community Colleges promoting scientific teaching while focusing on equity, assessment, and active learning.

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• Advisory Board Member • Completed week-long workshop focused on issues of scientific teaching, assessment, equity, diversity,

and active learning techniques during Summer 2010. • Attend several one-day workshops offered throughout the academic year • Partnered with a San Francisco State Graduate student to co-plan and co-teach classes during Fall

2010 and Fall 2012 • Participated in a learning community, called a Teaching Square, during Fall 2014 and Fall 2015 that

involved visiting each others’ classrooms and meeting regularly to discuss scientific teaching. American Society for Microbiology – Biology Scholars Assessment Fellow 2011-2012

• Attended three day intensive workshop focusing on assessment • Incorporated assessment formative and summative assessment techniques into community college

classroom • Utilized knowledge to present workshop on assessment and Student Learning Outcomes (SLO’s) to

CCSF Biology Department Faculty HONORS AND AWARDS

National Science Foundation Pre-doctoral Fellowship 2004 – 2007

Richard Fineberg Memorial Teaching Assistant Award 2005 Beta Cell Biology Consortium Retreat Travel Award 2003 & 2004 University of California Regents’ Fellowship, UCSF 2002 - 2003 Department of Nutritional Sciences Outstanding Senior, 2002 University of Arizona Summa Cum Laude 2002 Phi Beta Kappa 1999

PEER-REVIEWED PUBLICATIONS

Galli L, Santanta F, Szabo L, Ngo K, Apollon C, Burrus L. A toolbox for probing the palmitoylation, membrane trafficking, and transport of vertebrate WNT1. Developmental Biology. In revision (March 2018). Apollon, C. “Is the Data Dirty or Clean?”. National Center for Case Study Teaching in Science. University at Buffalo, State University of New York, 22 August 2017. Takeuchi JK, Lou X, Alexander JM, Sugizaki H, Delgado-Olguín P, Holloway AK, Mori AD, Wylie JN, Munson C, Zhu Y, Zhou YQ, Yeh RF, Henkelman RM, Harvey RP, Metzger D, Chambon P, Stainier DY, Pollard KS, Scott IC, Bruneau BG. Chromatin remodelling complex dosage modulates transcription factor function in heart development. Nat Commun. 2011 Feb 8;2:187 Munson C, Huisken J, Bit-Avragim N, Kuo T, Dong PD, Ober EA, Verkade H, Abdelilah-Seyfried S, Stainier DY. Regulation of neurocoel morphogenesis by Pard6 gamma b. Dev Biol. 2008 Dec 1;324(1):41-54. Epub 2008 Sep 9.

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Bit-Avragim N, Hellwig N, Rudolph F, Munson C, Stainier DY, Abdelilah-Seyfried S. Divergent polarization mechanisms during vertebrate epithelial development mediated by the Crumbs complex protein Nagie oko. J Cell Sci. 2008 Aug 1;121(Pt 15):2503-10. Epub 2008 Jul 15. Dong PD, Munson CA, Norton W, Crosnier C, Pan X, Gong Z, Neuman CJ & Stainier DY. Fgf10 regulates hepatopancreatic ductal system patterning and differentiation. Nature Genetics 2007 Mar;39(3):397-402.

Dodge JE, Munson C, List AF. KG-1 and KG-1a model the p15 CpG island methylation observed in acute myeloid leukemia patients. Leukemia Research. 2001 Oct;25(10):917-25. TEACHING WORKSHOPS, PRESENTATIONS, AND LECTURES

Presenter Post-Exercise Recovery Workshop March 2018

• Organized and led educational workshop on post-exercise recovery • Instruction on nutrition, hydration, and self-myofascial release

Panelist SFSU Postdoctoral Scientific Teaching Workshop April 2016

• Invited as a representative of the experiences teaching at a community college and small liberal arts school

Panelist UCSF Panel: Ask Faculty Members September 2014

• Launching Your Academic Job Search Series Presenter CCSF Professional Development Spring 2013 “Flipping, but not Flopping”

• A 5-part workshop series focusing on flipping the college classroom and active learning

Presenter Preparing Future Faculty (UCSF) 07/2010 “Using Science in the Media to Engage Undergraduates” • Part of a seminar series designed to aid UCSF staff and students

prepare for teaching

Workshop Presenter Expanding Your Horizons Skyline College - San Bruno, CA 2006 - 2009

Mills College - Oakland, CA 2005 San Francisco State University 2004, 2007-9

! A program encouraging middle and high school girls to pursue careers in math, science, and technology

! Designed and led hands-on workshops

Workshop Presenter Math, Science and CTE Conference (CCSF) 06/2009 and 06/2103 • Lead workshop on using zebrafish in K-12 classrooms

Presenter City College of San Francisco 10/2008 Biology Department Seminar Series

• “Zebrafishing for Genes”

Workshop Co-Presenter California Science Teachers Association Annual Conference 10/2006 “Scientific Inquiry and Genetics with Fruit Flies”

! Taught workshop on a series of lessons co-developed with a middle school science teacher

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Workshop Presenter San Francisco Unified School District Professional Development Day 03/2006 “Going zebrafishing”

! Taught hands-on workshop to middle and high school teachers ! Provided information on how to bring incorporate zebrafish into the

high school classroom

Panelist Preparing Future Faculty, UCSF Teaching Resources Panel 07/2005

! Served on a panel discussing how graduate students and postdoctoral fellows could gain teaching experience

RESEARCH PRESENTATIONS

Poster Presentation SABER West January 2017 “Let’s get graphing: Teaching Graphing Literacy on the first day of laboratory” • Discipline based education research project focusing on graph literacy West Coast Regional Developmental Biology Meeting 03/2007 Developmental Biology Program Retreat, UCSF 12/2006 “It’s Pard Time” International Zebrafish Conference – Madison, WI 06/2006 Munson CA, Huisken J, Kuo T, Horne-Badovinac S, Stainier DY. “Analysis of pard6gb and apico-basal polarity in neural tube lumen formation.” International Zebrafish Conference – Madison, WI 07/2004 Munson CA, Horne SA, Dong PD, Field HA, Stainier DY. “Asymmetric LPM Movement and Endodermal Organ Morphogenesis.” INTERESTS

Science & Biology Education

Scientific Teaching

Exercise Physiology

Incorporating current news stories and controversy into college science curriculum

Utilizing web-based media and technology to supplement course material ADDITIONAL SKILLS

Computer

! Computer Administrator for City College of San Francisco’s student laptops (for in-class use) ! Proficient in Microsoft Office, imaging, and publishing programs such as Adobe Illustrator,

Dreamweaver, and Microsoft Frontpage

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! Utilize Moodle-based software and Blackboard for creating technology-enhanced courses. ! Qualtrics survey software

Group Exercise Instructor ! YMCA of San Francisco 09/2017 - present ! UCSF Bakar Fitness Center – San Francisco, CA 10/2005 - present ! UCSF Millberry Fitness Center – San Francisco, CA 3/2003-present ! 24Hour Fitness – San Francisco, CA 9/2002-1/2004 ! University of Arizona Campus Recreation – Tucson, AZ 9/1999–7/2002 ! PiYo Certification Since 7/2017 ! Spinning® Certification (Star 3 Instructor) Since 11/2004 ! BodyPump® Certification Since 9/2007 ! CXWORX® Certification Since 1/2012 ! American Council on Exercise Fitness Nutrition Specialist Since 1/2016 ! American Council on Exercise Group Fitness Instructor Since 2001 ! American Council on Exercise Personal Trainer Since 2006 ! American Council on Exercise Health Coach Since 2010 ! CPR and First Aid Certification Since 1999 ! Regularly participate in professional development and continuing education coursework

Spanish ! Conversationally fluent ! Studied the Spanish language for 9+ years ! Participated in 5 week Spanish language immersion course in Guadalajara, Mexico