A type of soil bacterium that can transfer its own genes into plant cells and integrate into their genomes. Often used as a way to add new genes to crop plants.

One of several possible versions of a gene. Each one contains a distinct variation in its DNA sequence. For example, a “deleterious allele” is a form of a gene that leads to disease.

The chemical building block of proteins. During translation, different amino acids are strung together to form a chain that folds into a protein.

Microbes that look similar to bacteria but are actually more closely related to eukaryotes, such as humans. Archaea are single-celled organisms that don’t have a nucleus and can only be seen with a microscope. They’re found in many different habitats, and many of the first known examples were found in extreme environments.

An abundant type of microbe. These single-celled organisms are invisible to the naked eye, don’t have a nucleus, and can have many shapes. They’re found in all types of environments, from Arctic soil to inside the human body. Most bacteria are not harmful to human health, but certain pathogenic bacteria can cause illness.

The four “letters” of the genetic code (A, C, T, and G) are chemical groups called bases or nucleobases. A = adenine, C = cytosine, T = thymine, and G = guanine. Instead of thymine, RNA contains a base called uracil (U).

A type of genome editing that uses CRISPR components to directly change single DNA letters without making breaks in the DNA. Base editors are made by fusing (attaching) DNA-modifying enzymes to DNA-targeting Cas proteins like Cas9.

Different chemicals known as bases or nucleobases are found on each strand of DNA. Each base has a chemical attraction for a particular partner base, known as its complement. C matches up with G, while A pairs with T or U. These bonded genetic letters are called base pairs. Two strands of DNA can zip together to form a double-helix shape when complementary bases match up to form base pairs.

A type of disease caused by uncontrolled growth of cells. Cancerous cells may form clumps or masses known as tumors, and can spread to other parts of the body through a process known as metastasis.

Abbreviation of CRISPR-associated, may refer to genes (cas) or proteins (Cas) that protect bacteria and archaea from viral infection.

A protein derived from the CRISPR-Cas bacterial immune system that has been co-opted for genome engineering. Uses an RNA molecule as a guide to find a complementary DNA sequence. Once the target DNA is identified, Cas12 cuts both strands. Has been compared to “molecular scissors” or a “genetic scalpel.” In CRISPR immunity, cutting viral DNA prevents it from destroying the host cell. In genome engineering, cutting genomic DNA initiates a repair process that ends up making a change or “edit” to its sequence.

Note that Cpf1 (Cas12a), C2c1 (Cas12b), C2c3 (Cas12c), CasY (Cas12d), CasX (Cas12e), Cas14 (Cas12f), and CasPhi (Cas12j) are all Cas12 family members. Our illustration is based on Cas12a.

A protein derived from the CRISPR-Cas bacterial immune system that has been co-opted for genome engineering and CRISPR-based diagnostics. Uses an RNA molecule as a guide to find a complementary RNA sequence. Once the target RNA is identified, Cas13 cuts both the target and nearby, “collateral” RNA. In CRISPR immunity, cutting RNA like this is thought to cause “altruistic” cell suicide, preventing the spread of phages to neighboring cells. In genome engineering, an engineered version of Cas13 can be used to make a change or “edit” to targeted RNA sequences. In CRISPR-based diagnostics, Cas13’s collateral cutting activity can generate a fluorescent signal to indicate the presence of a pathogenic target RNA.

A protein derived from the CRISPR-Cas bacterial immune system that has been co-opted for genome engineering. Cas3 is a standalone nuclease-helicase that is recruited to targets by a separate, crRNA-guided complex called Cascade. Once Cascade identifies a complementary DNA target, it recruits Cas3 to move along the DNA, cutting as it goes. In CRISPR immunity, cutting phage DNA prevents phages from replicating and destroying the host cell. When used for genetic engineering, this approach can be used to delete thousands of base pairs around the target site. 

A protein derived from the CRISPR-Cas bacterial immune system that has been co-opted for genome engineering. Uses an RNA molecule as a guide to find a complementary DNA sequence. Once the target DNA is identified, Cas9 cuts both strands. Has been compared to “molecular scissors” or a “genetic scalpel.” In CRISPR immunity, cutting viral DNA prevents it from destroying the host cell. In genome engineering, cutting genomic DNA initiates a repair process that ends up making a change or “edit” to its sequence.

A protein complex derived from the CRISPR-Cas bacterial immune system that has been co-opted for genome engineering. Cascade is composed of multiple Cas proteins. The complex uses a crRNA as a guide to find a complementary DNA sequence. Once the target DNA is identified, Cascade recruits a separate nuclease-helicase called Cas3 to move along the DNA, cutting as it goes. In CRISPR immunity, cutting phage DNA prevents phages from replicating and it from destroying the host cell. When used for genetic engineering, this approach can be used to delete thousands of base pairs around the target site.

The basic unit of life. The number of cells in a living organism ranges from one (e.g. yeast) to quadrillions (e.g. blue whale). A cell is composed of four key macromolecules that allow it to function (protein, lipids, carbohydrates, and nucleic acids). Among other things, cells can build and break down molecules, move, grow, divide, and die.

The compact structure into which a cell's DNA is organized, held together by proteins. The genomes of different organisms are arranged into varying numbers of chromosomes, and human cells have 23 pairs.

The scientific term for cut or break apart. Typically refers to splitting apart a long polymeric molecule like DNA, RNA, or protein. For example, a nuclease like Cas9 can be directed to cleave DNA at a specific location.

Describes any two DNA or RNA sequences that can form a series of base pairs with each other. Each base forms a bond with a complementary partner. T (DNA) and U (RNA) bond with A, and C complements G. For example, in CRISPR immunity, the spacer sequence in a guide RNA is complementary to a sequence found in a viral genome. When the RNA bases pair with complementary DNA bases from an invading virus, the Cas9 protein will cut the target to stop the viral infection.

Pronounced “crisper.” An adaptive immune system found in bacteria and archaea, co-opted as a genome engineering tool. Acronym of “clustered regularly interspaced short palindromic repeats,” which refers to a section of the host genome containing alternating repetitive sequences and unique snippets of foreign DNA. CRISPR-associated surveillance proteins use these unique sequences as molecular mugshots as they seek out and destroy viral DNA to protect the cell.

During CRISPR immunity, the host cell generates crRNA molecules, each containing one spacer that is complementary to a portion of a viral genome. crRNAs guide CRISPR immune proteins to find and destroy matching invader sequences.

A technique that lets scientists see the effects of turning gene expression up or down with CRISPRa and CRISPRi. Instead of checking one gene at a time, a single CRISPR screen can provide information about thousands of different genes at a time.

CRISPRa stands for CRISPR activation and CRISPRi stands for CRISPR interference or inhibition. Both are methods for fine-tuning gene expression. If a gene were a car, CRISPRa is the gas pedal and CRISPRi is the brake. Using CRISPRa to activate a gene increases protein production. Using CRISPRi to “turn down” a gene reduces the number of protein products made from that gene.

Catalytically inactive, or “dead,” Cas9. This mutated version of the Cas9 protein cannot cut, but still binds tightly to a particular DNA sequence specified by the guide RNA. Can be used to physically block the process of transcription, turning off a specific gene, or to shuttle other proteins to a particular site in the genome.

Abbreviation of deoxyribonucleic acid, a long molecule that encodes the information needed for a cell to function or a virus to replicate. Forms a double-helix shape that resembles a twisted ladder. Different chemicals called bases, abbreviated as A, C, T, and G, are found on each side of the ladder, or strand. The bases have an attraction for each other, making A stick to T while C sticks to G. These rungs of the ladder are called base pairs. The sequence of these letters is called the genetic code.

When both strands of DNA are broken, two free ends are created. May be made intentionally by a tool such as Cas9. Cells repair their DNA to prevent cell death, sometimes changing the DNA sequence at the site of the break. Initiating or controlling this process with the intent to alter a DNA sequence is known as genome engineering.

A molecule, typically a protein, that causes or catalyzes a chemical change. Usually an enzyme’s name describes a molecule involved in the activity it performs and ends with the suffix -ase. For example, lactase is a well-known enzyme that breaks down lactose, a sugar found in milk. Cas9 is a nuclease, an enzyme that breaks apart the backbone of nucleic acids (RNA or DNA).

Refers to changes to a cell’s gene expression that do not involve altering its DNA code. Instead, the DNA and proteins that hold onto DNA are “tagged” with removable chemical signals. Epigenetic marks tell other proteins how to read the DNA, which parts to ignore, and which parts to transcribe into RNA.

Comparable to sticking a note that says “SKIP” onto a page of a book—a reader will ignore this page, but the book itself has not been changed.

A domain of organisms whose cells contain a nucleus and other organelles. Eukaryotes are often large and multicellular (e.g. elephants) but can also exist as microscopic, single cells (e.g. yeast). This category of life includes humans. Compare to prokaryotes (bacteria and archaea).

A product being made from a gene; can refer to either RNA or protein. When a gene is turned on, cellular machines “express” this by transcribing the DNA into RNA and/or translating the RNA into a chain of amino acids. For example, a “highly expressed” gene will have many RNA copies produced, and its protein product is likely to be abundant in the cell. CRISPRi and CRISPRa are methods for turning gene expression down or up, respectively.

A segment of DNA that encodes the information used to make a protein. Each gene is a set of instructions for making a particular molecular machine that helps a cell, organism, or virus function.

A mechanism for preferential inheritance of a particular DNA sequence. Usually, offspring have a semi-random chance of inheriting a given stretch of DNA from either parent. In a scientist-designed gene drive, a gene is engineered to have a 100% chance of being passed on. Gene drives can force the inheritance of a desirable trait through a population of organisms. For example, this approach could potentially make all mosquitoes incapable of transmitting the malaria parasite.

Delivering corrective DNA to human cells as a medical treatment. Certain diseases can be treated or even cured by adding a healthy DNA sequence into the genomes of particular cells. Scientists and doctors typically use a virus to shuttle genes into targeted cells or tissues, where the DNA is incorporated somewhere within the cells’ existing DNA. CRISPR genome editing is sometimes referred to as a gene therapy technique.

A genetically modified organism has had its DNA intentionally altered using scientific tools. Any organism can be engineered in this manner, including microbes, plants, and animals.

The entire DNA sequence of an organism or virus. The genome is essentially a huge set of instructions for making individual parts of a cell and directing how everything should run.

Intentionally altering the genetic code of a living organism. Can be done with ZFNs, TALENs, or CRISPR. These systems are used to create a double-strand break at a specific DNA site. When the cell repairs the break, the sequence is changed. Can be used to remove, change, or add DNA.

Repairing harmful DNA through a one-time genome editing procedure. Unlike taking a drug that will temporarily reduce long-term symptoms, altering a patient’s genetic code with the CRISPR-Cas9 “molecular scalpel” would permanently and directly reverse the cause of a genetic disease.

The study of the genome, all the DNA from a given organism. Involves a genome’s DNA sequence, organization and control of genes, molecules that interact with DNA, and how these different components affect the growth and function of cells.

The cells involved in sexual reproduction: eggs, sperm, and precursor cells that develop into eggs or sperm. The DNA in germ cells, including any mutations or intentional genetic edits, may be passed down to the next generation. In contrast, the genetic material in somatic cells (all the cells in the body except for germ cells) cannot be inherited by offspring. Note that genome editing in an early embryo is considered to be germline editing since any DNA changes will likely end up in all cells of the organism that is eventually born.

A two-piece molecule that Cas9 binds and uses to identify a complementary DNA sequence. Composed of the CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA). Cas9 uses the tracrRNA portion of the guide as a handle, while the crRNA spacer sequence directs the complex to a matching DNA sequence.

Scientists have also formed a version of the guide RNA that consists of a single molecule, the single-guide RNA (sgRNA).

A way for a cell to repair a break in its DNA by “patching” it with a piece of donor DNA. The donor DNA must contain similar sequences, or homology, to the broken DNA ends for it to be incorporated. HDR is a more precise repair pathway than non-homologous end joining. In genome engineering, a researcher designs and adds in the donor DNA, potentially allowing scientists to replace a disease-causing gene with a healthy copy.

Abbreviation for insertion or deletion. Refers to the random removal or addition of nucleotides from a DNA sequence. This can be enough to stop a gene from functioning (imagine removing a page from the middle of an instruction manual). Indels occur when DNA is broken and “sloppily” repaired by the cell in a process called