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Los Angeles County Science and Engineering Fair Research Plan for Experiments with Tissue, Cell Lines and/or Vertebrate Animals Certification Form 601-068

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Los Angeles County Science and Engineering Fair

Research Plan for Experiments with Tissue, Cell Lines and/or Vertebrate Animals

Certification Form 601-068    

Student(s) Name (Last, First)

Crusher, Beverly

Student Home Phone No.



Smallville MS

Site Science Fair Coordinator’s Name

Clark Kent

Site Coordinator Cell Phone Number


School Mailing Address

100 Main Street, Smallville, CA


Gotham City USD

Student Home Mailing Address

1 Farmdale Road, Smallville, CA

Student Email

Regenerating amputated human legs using Linkia sp. Sea Star tissue

Project Title


Problem: Will adding specialized genes from sea star tissue into humans who have lost their leg be able to regenerate new legs on humans?

Hypothesis: If regenerative genes extracted from DNA from Linckia sea stars is injected into humans at the wound site of an accidental amputation, then the lost limb will begin to grow.
1. Type of tissue, cell line, organ or organ parts, species, age and number of animals: Sea Star tissue from the arms of 10 Linkia sp.of sea stars will be used to locate genes for regeneration. Twenty humans between 21 – 40 years old who have voluntarily agreed to have the regeneration genes or a placebo (harmless liquid with no genes) inserted into leg tissue at the site of the missing limb.
2. Intended disposal of bio-hazardous materials, tissue, cell lines and/or vertebrate animals

(including post-research homes for live animals). Specify institutional procedures for management

of materials. Protocol # is not sufficient: The sea stars will have one of 5 arms amputated in their mid section and have the amputated piece used for tissue culture and gene discovery and manipulation. The sea stars have no brain to feel pain and will regenerate the missing portion of their arm over several weeks. The sea stars will be housed at the Gotham City Aquarium and released. The tissue and gene manipulation waste will be done in a “clean-room” facility at UC Gotham and any waste will be dealt with under state regulations for destruction.
3. Cite evidence of search for alternative to vertebrate animal use (i.e. use of bacteria, plants, invertebrates, In Vitro, etc.): I am using an invertebrate to extract the regeneration gene.
4. Bibliographic References (a minimum of 3 references, not exclusively Internet):

1. Edmondson, Charles Howard (1935). "Autotomy and Regeneration in Hawaiian Starfishes". Bernice P. Bishop Museum - Occasional Papers 11 (8): 6–12.

2. Tullis, Richard E.; Cheng, Thomas C. (1971). "The uptake of 14C by Stylifer linckiae (Mollusca: Prosobranchia) from its echinoderm host, Linckia multifora". Comparative Biochemistry and Physiology Part B: Comparative Biochemistry 40 (1): 109–10.

3. Aquarium Invertebrates: Sea Stars -- Linckia spp. By Rob Toonen, Ph.D.

Procedure/Research Techniques

(Attach additional pages if needed)
Provide a clear and detailed description/outline of proposed procedure, including equipment to be used, safety measures, description of humane treatment of vertebrate animals and/or specific use of tissue samples or cell lines, if applicable. Actual “procurement of and experimentation with” tissues, cell lines, and/or vertebrate animals is not allowed prior to the approval of this Research Plan by the Science Review Committee (SRC) of the Los Angeles County Science Fair.
Ten Linckia multifora Sea Stars from the Gotham City Aquarium touch tank will be selected for amputation of ½ an arm per animal. Amputations will be conducted under sterile conditions and the sea stars will be returned to holding tanks to recover and regenerate their arms. The removed tissue will be transported in sterile glass containers containing sea water, to UCG.
Under my mentor at UCG, the tissue will first be cleaned of parasites, wiped off with 5% bleach, crushed and soaked in buffer. The DNA in the meat should come off in the buffer.
After incubating the cells in a Luria nutrient broth, DNA extraction using a Phenol/Chloroform method will be conducted and the DNA will be amplified via PCR. I will use DNA probes known from previous research to identify possible genes involved in regeneration. Current studies suggest that the use of macrophage-derived therapeutic molecules could actually help humans regrow limbs right in the emergency room.
Voluntary participants for the project will be selected from recent trauma victims at the Gotham City Hospital and the Smallville Community Hospital, since the Greater Gotham area is subject to frequent disasters. Participants will fill out consent forms and have gene therapy containing the Linckia-derived molecules applied by doctors, under our stated protocol. I will follow their progress, record any side effects and measure any tissue growth for a period of 3 months. Results will be collected in data tables and graphs. Statistical analysis will be performed, including multi-variate analyses and data from CAT scans.

Research Plan/Tissues and/or Vertebrates

Revised- May 2013

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