Author Profile

Dr. Frank J. Slack is an internationally acclaimed pioneer in genetics, a foundational figure in the small RNA revolution, and a leading authority at the frontier of functional genomics. As the Shields Warren Mallinckrodt Professor of Medical Research and Pathology at Harvard Medical School and the Director of the Institute for RNA Medicine at Beth Israel Deaconess Medical Center (BIDMC), Dr. Slack brings decades of transformative, Nobel-adjacent research directly to the pages of Principles of Gene Manipulation and Genomics. His work sits at the very intersection of genetic engineering, disease therapeutic development, and large-scale genome analysis. Dr. Slack’s academic journey began with a B.Sc. from the University of Cape Town in South Africa, followed by a Ph.D. in molecular biology from the Tufts University School of Medicine. He cemented his place in scientific history during his postdoctoral fellowship in the laboratory of Gary Ruvkunat Harvard Medical School. There, Dr. Slack co-discovered let-7—only the second microRNA ever identified and the first-ever discovered in humans. This milestone fundamentally shifted the global understanding of gene regulation, proving that tiny, non-coding pieces of RNA hold the master switches for complex cellular systems. Throughout his highly decorated career—spanning leadership roles as a program director at the Yale Center for RNA Science and Medicine to his current chair at Harvard—Dr. Slack has continually pushed the boundaries of what is possible in genomic manipulation.

About The Book

Principles of Gene Manipulation and Genomics stands as the definitive blueprint for understanding, navigating, and mastering the genetic revolution.From the foundational mechanics of recombinant DNA technology to the artificial synthesis of entire genomes, this book offers an unparalleled, rigorous exploration of how we isolate, analyze, and engineer genetic material.This text is not merely a collection of facts; it is a conceptual roadmap designed for advanced undergraduates, graduate students, and researchers. It systematically dismantles complex methodologies into elegant, understandable frameworks. What sets this edition apart is its relentless focus on application. Principles of Gene Manipulation and Genomics does not just explain how a technology works; it illustrates why it matters. Through real-world case studies and industrial insights, the book highlights the pivotal role of genomics:Precision Medicine: How gene therapies, mRNA platforms, and personalized oncology are turning once-fatal genetic disorders into treatable conditions.Agricultural Sustainability: The development of climate-resilient crops, enhanced bio-fortification, and engineered microbes designed to secure the global food supply.The Bioeconomy: The deployment of engineered enzymes and metabolic pathways for green chemistry, biofuel production, and bioremediation. Featuring crisp, high-resolution schematics, lucid pedagogical features, and up-to-date references to pioneering literature, Principles of Gene Manipulation and Genomics remains the ultimate companion for anyone serious about molecular biology. Whether you are a student preparing for a career at the bench, a researcher looking to expand your technological toolkit, or a biotech professional navigating the frontier of innovation, this book provides the foundational knowledge and visionary insight required to shape the future of biological science.

Table of Contents

Preface
. GENE MANIPULATION IN THE POST-GENOMICS ERA
. Introduction
. Gene manipulation involves the creation and cloning of recombinant DNA
.
. BASIC TECHNIQUES
. Introduction
. Blotting is used to transfer nucleic acids from gels to membranes for further analysis
. Western blotting is used to transfer proteins from acrylamide gels to membranes
. The basic PCR is not efficient at amplifying long DNA fragments
.
. CUTTING AND JOINING DNA MOLECULES
. Cutting DNA molecules
. Joining DNA molecules
. DNA molecules can be joined without DNA ligase
.
. BASIC BIOLOGY OF PLASMID AND PHAGE VECTORS
. Plasmid biology and simple plasmid vectors
. The host range of plasmids is determined by the replication proteins that they encode
. Bacteriophage l
. DNA cloning with single-stranded DNA vectors
.
. COSMIDS, PHASMIDS, AND OTHER ADVANCED VECTORS
. Vectors for cloning large fragments of DNA
. Specialist-purpose vectors
. Proteins that are synthesized with signal sequences are exported from the cell
.


. GENE-CLONING STRATEGIES
. Introduction
. Genomic DNA libraries are generated by fragmenting the genome and cloning overlapping fragments in vectors
. The PCR can be used as an alternative to genomic DNA cloning
. Complementary DNA (cDNA) libraries are generated by the reverse transcription of mRNA
. The PCR can be used as an alternative to cDNA cloning
. Many different strategies are available for library screening
.
. SEQUENCING GENES AND SHORT STRETCHES OF DNA
. The commonest method of DNA sequencing is Sanger sequencing (also known as chain-terminator or dideoxy sequencing)
. Alternatives to Sanger sequencing have been developed and are particularly useful for resequencing of DNA
.
. CHANGING GENES: SITE-DIRECTED MUTAGENESIS AND PROTEIN ENGINEERING
. Introduction
. Primer extension (the single-primer method) is a simple method for site-directed mutation
. The PCR can be used for site-directed mutagenesis
. Phage display can be used to facilitate the selection of mutant peptides
. Protein engineering
. A number of different methods of gene shuffling have been developed
.
. BIOINFORMATICS
. Introduction
. Databases are required to store and cross-reference large biological datasets
. Sequence analysis is based on alignment scores
. Sequence analysis of genomic DNA involves the de novo identification of genes and other features
. Several in silico methods are available for the functional annotation of genes
.


. Caution must be exercised when using purely in silico methods to annotate genomes
. Sequencing also provides new data for molecular phylogenetics
.
. 10. MAPPING AND SEQUENCING GENOMES
. Introduction
. Physical mapping is limited by the cloning process
. Optical mapping is undertaken on single DNA molecules
. Sequencing genomes
. The quality of genome-sequence data needs to be determined
.
. 11. COMPARATIVE GENOMICS
. Introduction
. The formation of orthologs and paralogs are key steps in gene evolution
. Comparative genomics of bacteria
. Comparative genomics can be used to analyze physiological phenomena
. Mitochondrial genomes exhibit an amazing structural diversity
. Horizontal gene transfer has been detected in mitochondrial genomes
. Comparative genomics of eukaryotes
.
. 12. LARGE-SCALE MUTAGENESIS AND INTERFERENCE
. Introduction
. Genome-wide gene targeting is the systematic approach to large-scale mutagenesis
. Genome-wide random mutagenesis is a strategy applicable to all organisms
. Transposon mutagenesis in plants can be achieved using endogenous or heterologous transposons
. Insertional mutagenesis in invertebrates
. Libraries of knock-down phenocopies can be created by RNA interference
.

BIBLIOGRAPHY
INDEX