RecombinantDNA: History of the Controversy

Charles L. Vigue WilliamG. Stanziale

W HENTHE from two differentorganisms are recombinantDNA molecules are discussed in Cohen joined, the resultant and Sinsheimer(1977). moleculeis referredto as recombi- (1975), Grobstein(1977) Downloaded from http://online.ucpress.edu/abt/article-pdf/41/8/480/37474/4446723.pdf by guest on 29 September 2021 nant DNA. The DNAs involvedcan be from organisms as close as the same or as distantas different Biohazards kingdoms. Recombinant DNA and the technology The insertionof DNA fromone lifeform into eitherthe involved in its construction are controversial.Several same or a differentspecies could produce cellularmeta- books have been publishedabout recombinant DNA, its bolicchange(s) resulting in undesirablecharacteristics in history and controversy. Among the more popular the recipientorganism. The resultantchimera, if a bacte- books are RecombinantDNA: The Untold Story, by rium,would reproduceat a rapidrate and possess bio- John Lear;Biohazard, by MichaelRogers; The Ultimate chemicalproperties that may not be knownbeforehand. Experiment,by Nicholas Wade; and Playing God, by DNA from one bacterialspecies or strain could be June Goodfield.This paperreviews for biologyteachers into the genic complementof anotherbacterium, the history and controversy surroundingthe recombi- spliced or unknowinglyproducing a chimerathat is nant DNA debate. knowingly highly infectious or cancer-causing,or one that pos- The process of constructing a recombinant DNA resistanceto availableantibacterial drugs used for molecule usuallyinvolves combininga specific DNA of sesses The accidental escape of such a chimera interest from one species, the donor, with that of an treatment. could resultin creationof a new disease or propagation appropriatevector (, bacteriophageor ). of increasing populations of drug-resistant strains The vector carries the donor DNA into an appropriate naturalsexual matingand transductionof bac- host species, the recipient,such as Escherichiacoli, but (through terial ). Laboratorycontrol is of paramount it does not combine with the host. The ultimateaim is importance and must include frequent monitoringof twofold-to amplifythe DNA of interest in a unicellular procedures.This can be readilyachieved because much life form so it may be more easily studied,and to make of the technologyof biohazardcontrol is available,hav- the DNA functionin the productionof a specific RNA, ing been developedover a periodof some thirtyyears at , or protein.Various methods used to construct the Fort Detrick Biological Laboratories,Frederick, and Charles L. Vigue is assistant Maryland.This technology is highly sophisticated professorof ,and Wil- includes methods for intense monitoringof technical liamG. Stanzialeis professor proceduresand personnel. of biologyand chairman of the biologydepartmentandmedi-re c altechnology program direc- Social Implications tor at Saint Joseph College, 1678 Asylum Avenue, West The social benefitsassociated withrecombinant DNA Hartford,Connecticut 06177 1=. and technology are unprecedentedin human Vigue received his B.A. and the resolution M.S.degrees in zoologyfrom the Universityof Mainein 1968and 1970 history.This scientificeffort could lead to respectively;he was awarded his doctorate in by North of many medicaland social problemsthrough commer- CarolinaState Universityin 1973.He has taughtat Essex Community cial applicationof this knowledge for manufacturing College(Baltimore, Maryland) and Johns HopkinsUniversity. He is a and memberof AAAS, AIBS,and Sigma Xi. Stanzialereceived his B.S. greaterquantities and diversityof pharmaceuticals degreefrom Wilkes College (Wilkes-Barre, Pennsylvania) and his M.S. other medicallyrelated chemicals, developmentof new and Ph. D. degreesfrom Catholic University of America(Washington, cereal grains for humanand animalconsumption, pro- D.C.) in 1955 and 1960 respectively. He served as a research of available microbiologistat the U.S. Departmentof Defense Laboratories,Ft. duction of new and improvement Detrick,Maryland, from 1952through 1961, and was a facultyfellow vaccines, creationof additionalnitrogen-fixing bacteria and visitingscientist in 1969-70at the NASA MannedSpacecraft capable of livingin a diversityof plantspecies to signifi- Center in Houston, Texas. Stanzialeis a memberof the Society of of soil, replacement IndustrialMicrobiologists and AIBS. cantly increase naturalfertilization

480 THE AMERICANBIOLOGY TEACHER,VOLUME 41, NO. 8, NOVEMBER1979 of defectivehuman genes to eradicategenetic diseases, Nucleic Acids held on June 11-15,1973, in New Hamp- and precise mapping of human genes to achieve an ton, New Hampshire.At this meeting, of understandingof the molecularbasis of humandisease, the Universityof CaliforniaMedical Center presented especiallycancer. the results of his recombinant DNA experiments in The dawningof this era of recombinantDNA could which he collaboratedwith Stanley Cohen of the Stan- also be demonstratedin an awesome manner, by the ford UniversityMedical Center. It was suggested to the intentionaluse of this knowledge by some humans to Conference Co-chairpersons, of the controlother groupsor races of humanity.The profound National Institutefor Allergicand InfectiousDiseases implicationsfor abusive applicationshould be a special and Dieter Soll of Yale University,that they allowfor a concern in considering the probable application of short discussion of the potentialrisks of recombinant recombinantDNA for genetic engineeringof humans. DNA experimentation. Responsibilityfor technicalliteracy must be assumed The next morning the Conference addressed the by the publicand responsibilityfor educatingthe public potential risks of recombinantDNA experimentation. must be assumed by the scientists. The assessment of The majorityof participantsvoted to address a letter of this technologymust be accomplishedon a world-wide concern about recombinantDNA technology to the basis, and ultimately internationalcontrol must be Presidentsof the NationalAcademy of Sciences (NAS) achieved. and the Instituteof ,and to Science, the publi- Downloaded from http://online.ucpress.edu/abt/article-pdf/41/8/480/37474/4446723.pdf by guest on 29 September 2021 cationof the AmericanAssociation for the Advancement Evolutionof the Controversy of Science (AAAS).The letter, publishedin Science on The controversy over recombinant DNA had its September 21, 1973, said that new recombinantDNA beginningin late June, 1971, at a Cold SpringHarbor technologymay create "hybridplasmids or viruseswith Laboratory(Cold Spring Harbor, New York) tumor biological activity of unpredictable nature. . . Certain virus workshop.The workshopwas attended by Janet such hybridmolecules may prove hazardousto labora- tory workersand to the public.Although no hazardhas Mertz,a graduatestudent in the laboratoryof PaulBerg, yet been established,prudence suggests that the poten- Professorof Biochemistryat StanfordUniversity Medi- tial hazards be seriously considered."The letter also cal Center. Mertz told Robert Pollack, a Cold Spring suggested that a committee be establishedto consider Harbor Laboratorystaff member, of one of Berg's upcoming experiments designed to recombine Simian the problem and to recommend certain actions or Virus-40(SV-40) DNA with phage lambdaDNA and to guidelines. insertthe hybridmolecule into E. coli. SV-40is knownto cause tumorsin monkeysand certain other primates and Developmentof NIH Guidelines to cause the transformationof normal human tissue The NAS respondedto the letterby askingPaul Berg culturecells into cells with tumor-likecharacteristics. to form a NAS committee to pursue the problemsof Because E. coli is a naturalinhabitant of the human what to do about the recombinantDNA controversy. large intestine, Pollackknew of the potentialdanger- The BergCommittee, officially named the Committeeon that an E. coli containingSV-40 DNA may cause tumors Recombinant DNA Molecules, consisted of several in humans.Thus, Pollacktelephoned on June prominentscientists. 28, 1971to discuss the possibledangers. Although Berg At its firstmeeting in April,1974, at the Massachusetts understoodthe infectivityof SV-40,he did not consider Instituteof Technology,the Berg Committeedecided to his experimentsoverly dangerous. call for an internationalconference sponsored by the By early 1972, severalscientists had expressed con- NAS to be held at Asilomar in February,1975. The cern over Berg's experiments.As Berg was unable to mission of the conference would be to determinethe assign a zero risk to his experiments, he decided to degree of riskof variousrecombinant DNA experiments. abandonhis plans.Subsequently, he informedPollack of In the meantime,the Committeecalled for a moratorium his decision and asked Pollackto organizea meetingto on recombinantDNA research that may extend onco- discuss the safety of recombinantDNA research. gentic DNA molecules or antibiotic resistant DNA The meeting, the Conference on the Safety and molecules to species not naturallyharboring them. An Hazards of Tumor , was convened on January open letter from the Committee was published in 22, 1973, at the AsilomarConference Center at Pacific Science on July26, 1974,in Natureon July19, 1974,and Grove, Californiaand it lasted for three days. At that in the July issue of the Proceedings of the National meeting,Berg calledon the NationalInstitutes of Health Academyof Sciences. The lettersaid that certain recom- (NIH)to conduct epidemiologicalexperiments to deter- binantDNA experiments"would... resultin the creation mine the risk of oncogenic viruses to individuals.The of novel types of infectiousDNA elementswhose biologi- NIHI,however, did not heed the request. cal properties cannot be completely predicted in The debate among the scientists as to the safety of advance. There is serious concern that some of these recombinantDNA experimentscontinued to grow and artificialrecombinant DNA moleculeswould prove bio- surfaced publicly during the Gordon Conference on logicallyhazardous."

RECOMBINANTDNA 481 The letter also asked NIH to establish "an advisory committee charged with (i) assessing an experimental TABLE1. VariousRecombinant DNA Experimentsand RequiredContainment as Specifiedby the InitialGuidelines programto evaluatethe potentialand ecological hazards of the above types of recombinantDNA molecules, (ii) developingprocedures which will minimize the spreadof DNA Into E. coli Containment such molecules within human and other populations, Nonembryonicprimate tissue P3 EK3or P4 EK2 and (iii)devising guidelines to be followedby investiga- Embryonictissue or germ line cells tors working with potentiallyhazardous recombinant from primates P3 EK2 DNA molecules." Nonprimatemammalian tissue P3 EK2 One hundredfifty-five scientists, lawyers,and media Birds P3 EK2 Cold blooded vertebrates(non- representativesattended the four-dayNAS International embryonictissue) P2 EK2 Conference on RecombinantDNA molecules which Lowereukaryotes P2 EK1 opened on February24, 1975,and lasted for fourdays. Prokaryotesthat exchange genes The Conference covered five topics-ecology of plas- with E. coli- mids and enteric bacteria,molecular biology of proka- Non-pathoqens P1 EK1 Low-riskpathogens P2 EKI ryote plasmids and their use for molecular , Moderate-riskpathogens P2 EK2 synthetic recombinantsinvolving animal virus DNAs, Prokaryotesthat do not exchange Downloaded from http://online.ucpress.edu/abt/article-pdf/41/8/480/37474/4446723.pdf by guest on 29 September 2021 syntheticrecombinants involving eukaryotic DNAs, and genes with E. coli- ethical and legal concerns. Moderate-riskpathogens P2 EK2 Animalvirus P4 EK2or P3 EK3 An organizingcommittee for one of the topics sug- Plantvirus P3 EKI or P2 EK2 gested possible guidelinesfor recombinantDNA experi- Plasmidor phage naturallyexchang- ments. The same group suggested banning certain ing genes withE. coli P1 EK1 high-riskexperiments such as introducinga gene for a DNAs from highlypathogenic organ- deadlytoxin into E. coli. Prioritywould be givento devel- isms and oncogenic organismsof moderaterisk Banned oping a weakenedE. coli that wouldhave extreme diffi- DNAs containinggenes for toxins Banned culty surviving if it escaped from the laboratory DNAs of plantpathogens which may environment. increase virulenceor host range Banned DNAs transferringdrug resistance to Several scientistswere concerned that the guidelines organismsnot naturallyacquiring might result in legislation. Discussion ensued as to it Banned whetheror not thereshould be a controlon recombinant that and the DNA research. Some felt the guidelines From ,Human Genetics, and Cell Biology: moratoriumwere infringementson theiracademic free- Evolution of TechnologicalIssues, DNA RecombinantMolecule dom and freedom of inquiry.The majorityof the dis- Research,Supplemental Report II, Report preparedfor the theirminds Subcommitteeon Science,Research and Technology of theCommit- senters againstthe guidelineschanged when tee on Scienceand Technology, U.S. Houseof Representatives. RichardDworkin, a Professor of Law, explained that heavy fines may be leviedagainst scientists if they were found negligentin any deaths or illnessesresulting from use the recommendationsof the AsilomarConference their work. as interimguidelines. The second meeting of the NIH Committeewas on On the final day of the Conference the participants May12 and 13, 1975.A sub-committeechaired by David adopteda statementthat allowedthe recombinantDNA Hogness of Stanford Universityaccepted the task of research to continue but only under adequate safe- developingmore specificNIH guidelines. Biological con- guards includingboth physical and biologicalcontain- tainment(disarmed hosts) andthe specificityof the crite- ment. The guidelines proposed by the Organizing ria to be used in selecting physicalcontainment were Committee includedfour risk categories rangingfrom discussed. minimalto high. The degree of physicaland biological At the thirdmeeting held at Woods Hole, Massachu- containmentincreased as the potentialrisk increased. setts in July, 1975, the "Hogness" guidelines were The conference participantsvoted to includea ban on revised.At this meeting,controversy developed over the experimentsinvolving DNAs of pathogenicorganisms. degree of containmentnecessary for recombinantDNA In addition,the conferees voted to extend the ban until research. Some felt that the 'Woods Hole" guidelines suitablevectors were developed. were too lax, and others felt they were too strict. On February 26, 1974 the new NIH Recombinant Because so much controversyevolved at the Woods DNA Molecule ProgramAdvisory Committee, estab- Hole meeting,Chairman Stetten decidedto circulatethe lished on October 7, 1975 and chaired by NIH Deputy guidelines within the scientific community.The many Director DeWitt Stetten, Jr., met to discuss the state- commentsreceived suggested that anothermeeting was ments made at the AsilomarConference and to draftits necessary. ElizabethKutter of EvergreenState College, own specificguidelines. The NIHCommittee decided to Olympia,Washington, accepted the task of preparing

482 THE AMERICANBIOLOGY TEACHER,VOLUME 41, NO. 8, NOVEMBER1979 yet another set of revised guidelines.On December 4 and 5, 1975,the Committeemet at LaJolla,California to TABLE2. VariousRecombinant DNA Experimentsand compare the "Hogness,""Woods Hole,"and "Kutter" RequiredContainment as Specifiedby the RevisedGuidelines guidelinesand to formulatea finalset of guidelines. The guidelineswere discussed at the NIHmeeting on DNA Into E. Coli Containment February9 and 10, 1976, in Washington,D.C. The final PrimateDNA P2 EK2 guidelines were published in the Federal Register on Nonprimatemammals P2 EK2 June 7, 1976. Birds P2 EK2or P3 EK1 Cold-bloodedvertebrates P2 EKi or P1 EK2 The NIHGuidelines Lowereukaryotes (species producing polypeptidetoxins moderately The finalguidelines governing research at NIHlabora- active in vertebrates) P3 EK2 tories and of its grantees and contractors are stricter Lowereukaryotes (other) P2 EKI or P1 EK2 Plants(not producingtoxins) P2 EKi or P1 EK2 than those recommendedby the confereesat Asilomar. Prokaryotesexchanging genes with These guidelinesuse two containmentsystems-one for E. coli UsuallyExempt physical(P) containmentand one for modifiedand dis- (If not exempt) P1 EKI E. K 12 EKlevels Prokaryotesnot exchanginggenes armed coli strain (EK).There are three Downloaded from http://online.ucpress.edu/abt/article-pdf/41/8/480/37474/4446723.pdf by guest on 29 September 2021 and four P levels of containment: with E. coli UsuallyP2 EKI or P1 EK2 EK 1-The standardE. coli K 12 can be used. Virusesof Eukaryotes Varies from P1 EK 2-E. coliK 12with a chanceof survival of less than one EK1to P2 EK2or in 108must be used. P3 EKI depending EK 3-This is the sameas EK2 onlythe survival has been on DNA used testedin livingorganisms. P 1-Only standardmicrobiological procedure need be used. P 2-Extra precautionsto preventaerosol dispersion are required. TABLE3. ExperimentsExempt from the RevisedGuidelines P 3-Negativeair pressure inside the laboratory to prevent and Prohibited airescape is required. P 4-Air locks, protectiveclothing and showeringare required. Experiment Classification The least hazardousexperiments would combine EK 1 Formationof recombinantDNA from highlypathogenic organ- with P 1, and the most hazardouswould combine EK 3 isms Banned with P 4 containment.Experiments classified under the Formationof recombinantDNA variouscontainment systems are outlinedin table 1. forminghighly active toxins active On September 27, 1977, after several months of in vertebrates Banned review, the NIH RecombinantDNA MoleculeProgram Formationof recombinantDNA whichconfers increasedvirulence Advisory Committee publishedproposed revisions to or host range of a plant pathogen Banned the initialguidelines. The final revised NIH Guidelines Transferof a drug resistancefactor were publishedin December 1978. The new guidelines to an organismnot acquiringit ease the restrictionsof the previousones (table2). Cer- naturally Banned tain experiments are exempt from the guidelinesand Use of DNAs consistingentirely of DNA segments from a single non- others are prohibited(table 3). Moreemphasis is placed chromosomaeor viralDNA source Exempt on local biosafetycommittees. DNAs that consist entirelyof DNA Althoughthe NIH Guidelinesspecify the practicesto from a prokaryotichost when be used forconstructing and handling recombinant DNA propagatedin that host Exempt moleculesand organismscontaining recombinant DNA molecules, ultimateresponsibility for biosafety resides thesis of toxins potent for vertebrates.The guidelines with the institutionwhere this researchis conducted.At also prohibitthe deliberatecreation of a plantpathogen each institution,a Biosafety Committee, a Biological with increased virulence and host range. Large-scale Safety Officer and the principalinvestigator of the experiments(more than 10 litersof culture)with organ- research constitute a team to ensure adherence to the isms containingrecombinant DNAs are also excluded guidelines,but final accountabilityfor biosafetyappar- unless the DNAs are rigorouslycharacterized and the ently rests with the head of the institution. harmlessness of the recombination has been The guidelineseffectively prohibit specific experimen- established. tal procedures that could have serious social implica- Exceptionsto these prohibitionsmust be approvedby tions. The formation of recombinant DNAs from the Directorof the NationalInstitutes of Healthwith the pathogenicorganisms is prohibited,as wellas formation advice of a nationalRecombinant DNA AdvisoryCom- of recombinantDNAs containinggenes directingbiosyn- (Continued on p. 491)

RECOMBINANTDNA 483 Microbiology Labs References ...from p. 486 DEAN, D.S. 1970. Preservice preparation of college biology References teachers: a search for a better way. Washington, D.C.: Commissionon UndergraduateEducation in the Biolog- BURKE, D.D. 1976. Mini-Investigativelabs in microbiology for ical Sciences. non-science students. AIBS Ed. Reu. 5-1. NAPELL,S. 1974. TA. Handbook. Graduate Assistants' HOLT, C.E., ABRAMOFF, P., WILCOX, L.D. JR., and TeachingProgram, University of California,Berkeley. ABELL, D.J. 1969. Investigative laboratory programs in NORBERG,A. 1975.Individualizing instruction in largeunder- biology. Bio-Science 19:1104. graduatebiology laboratories II: computers and investiga- THORNTON, J., ed., 1971. The laboratory: a place to investi- tion. American Biology Teacher 37(8):470. gate. (Publication number 33). The Commission on Under- VON BLUM,R. 1973.An innovativeapproach to laboratory graduate Education in the Biological Sciences, Washington, instruction. AIBS Educational Review 2(2):17. D.C. . 1975. Individualizing instruction in large under- VON BLUM, R.C. 1973. An innovative approach to laboratory graduate biology laboratories I: development of the instruction. AIBS Ed. Rev. 2:1. model. American Biology Teacher 37(8):467. Recombinant DNA Downloaded from http://online.ucpress.edu/abt/article-pdf/41/8/480/37474/4446723.pdf by guest on 29 September 2021 ...from p. 483 mittee. Very significantand noteworthyis the fact that Teaching Assistants appropriateopportunity must be provided for public ...from p. 479 comment. However,the guidelinesapply only to recom- binantDNA researchperformed at institutionsreceiving dinatorsfelt unpreparedfor some of theirduties, such as NIH funds for research and research conducted at the moderating group discussions and making in-class NIH. Apparently,adherence to the guidelinesis volun- observationsof TAs. The changessuggested here are(1) tary for other institutions. to reduce the teachingor programduties of the coordi- nators,and (2) to preparecoordinators through orienta- tion sessions with TA trainingexperts. References Conclusion COHEN,S. 1975.The manipulationof genes. Sci. Am. 233:24. COOKE,R. 1977.Improving on nature.New York:Dameter The TA trainingprogram in Biology 1 at Berkeley Press. revealed three importantpoints. First, a TA training GLASS, B. 1977. The scientist: trustee for humanity.Bio- programfor a large undergraduatescience course can Science 27:277. be implemented successfully by experienced TAs. GOODFIELD,J. 1977.Playing God: genetic engineeringand a course TA trainingprogram can establish manipulationof life. New York:Random House. Second, GROBSTEIN,C. 1977. The recombinantDNA debate. Sci. good working relationshipswith other TAs as well as Am. 237:22. providea forum where teachingdifficulties may be dis- HOWARD,T. and RIFKIN,J. 1977. Who shouldplay God: cussed and solutions sought. Third,student ratingsof New York:Delacorte Press. TAs seem to improve as a result of such a training LEAR,J. 1978. RecombinantDNA: the untold story. New York:Crown Publishers, Inc. program. PAOLETTI,R. (ed). 1974. Selected readings:genetic engi- The followingaspects of the modelpresented here are neeringand .New York:Arno Press. most usefulfor establishinga trainingprogram for teach- ROGERS,M. 1977. Biohazard.New York:Alfred A. Knopf, ing assistants in undergraduatebiology courses: Inc. 1. The program should be designed by faculty SINSHEIMER,R. 1977. RecombinantDNA. Ann. Rev. Bio- and TAs, and largelyby the TAs chem. 46:415. members implemented UNITEDSTATES DEPARTMENTOF HEALTH,EDUCA- themselves,who haveexperience in teachingthe course. TION, AND WELFARE,NATIONAL INSTITUTES OF 2. In addition to pre-service sessions, ongoing in- HEALTH.1976. RecombinantDNA research:guidelines. service sessions and evaluationsshould be conducted. FederalRegister, 7 July 1976. 3. Each TA should have his/her own coordinatorto -__ . 1978. Guidelines for research involving recombinant DNA molecules. Federal Register, 22 consult. December 1978. 4. The TA trainingprogram should be integratedwith WADE, N. 1977. The ultimateexperiment: man-made evo- the course. lution.New York:Walker.

MICROBIOLOGYLABS 491