Crispr Engineering
Crispr Engineering - Related Videos
06:12
Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex
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Cited by 24 •
2018
Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system.
02:53
CRISPR and crRNAs
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2020
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea. The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
08:20
Genome Engineering of Primary Human B Cells Using CRISPR/Cas9
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Cited by 13 •
2020
Here we provide a detailed, step-by-step protocol for CRISPR/Cas9-based genome engineering of primary human B cells for gene knockout (KO) and knock-in (KI) to study biological functions of genes in B cells and the development of B-cell therapeutics.
06:33
Production of Human CRISPR-Engineered CAR-T Cells
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Cited by 19 •
2021
Here, we present a protocol for gene editing in primary human T cells using CRISPR Cas Technology to modify CAR-T cells.
11:53
Substrate Generation for Endonucleases of CRISPR/Cas Systems
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Cited by 4 •
2012
CRISPR/Cas systems mediate adaptive immunity in Bacteria and Archaea. Many Cas proteins are proposed to act as endoribonucleases acting on crRNA precursors of varying length. Here we illustrate three different approaches to generate pre-crRNA substrates for the biochemical analysis of Cas endonuclease activity.
FAQs
Target recognition depends on sequence matching between the guide RNA and the chosen DNA or RNA region. The guide positions the Cas protein at that site, allowing the system to cleave the target or carry out a programmed modification. This targeting step connects a specific genetic sequence with a measurable biological function, making it useful for testing causal relationships.
CRISPR engineering can be configured around cleavage followed by template-directed repair, base editing, or related modification strategies. These approaches differ in how the targeted genetic material is changed after the guide RNA and Cas protein identify the sequence. Selecting among them helps align the genetic intervention with the biological question, including studies of host functions and infection-related mechanisms.
The target type determines which genetic material is examined. DNA-directed systems can investigate host or pathogen genetic sequences, whereas RNA-directed systems focus on matching RNA sequences. This distinction allows researchers to connect the CRISPR engineering strategy with different questions about immune responses, pathogen replication, or mechanisms that contribute to immune evasion.
A conceptual workflow begins by selecting a host or pathogen sequence linked to the biological question, then designing a matching guide RNA and pairing it with an appropriate Cas system. Researchers next choose cleavage, template-directed repair, base editing, or a related approach, and examine how the resulting genetic change affects the function under study.
Researchers can target selected host genes and examine how their alteration changes immune responses. This approach links particular genetic factors to the behavior of immune systems, helping distinguish components that promote, limit, or otherwise control defense. In infection studies, such findings can reveal host processes that influence pathogen growth or the effectiveness of antimicrobial strategies.
CRISPR engineering can create immune cells with selected genetic changes or produce disease models that reproduce specific host or pathogen functions. These systems help researchers investigate mechanisms, evaluate therapeutic development, and test antimicrobial strategies in a controlled biological context. The resulting models connect targeted genetic manipulation with measurable consequences for immunity and infection.