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Generation of Efficient Knock-in Mouse and Human Pluripotent Stem Cells Using CRISPR-Cas9
- Source :
- Methods in molecular biology (Clifton, N.J.). 2320
- Publication Year :
- 2021
-
Abstract
- A knock-in can generate fluorescent or Cre-reporter under the control of an endogenous promoter. It also generates knock-out or tagged-protein with fluorescent protein and short tags for tracking and purification. Recent advances in genome editing with clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9) significantly increased the efficiencies of making knock-in cells. Here we describe the detailed protocols of generating knock-in mouse and human pluripotent stem cells (PSCs) by electroporation and lipofection, respectively.
- Subjects :
- Gene Editing
Induced Pluripotent Stem Cells
Drug Resistance
Recombinational DNA Repair
Clone Cells
Culture Media
Mice, Inbred C57BL
Mice
Electroporation
Genes, Reporter
Animals
Humans
Puromycin
Gene Knock-In Techniques
CRISPR-Cas Systems
Cells, Cultured
Embryonic Stem Cells
DNA Primers
RNA, Guide, Kinetoplastida
Subjects
Details
- ISSN :
- 19406029
- Volume :
- 2320
- Database :
- OpenAIRE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Accession number :
- edsair.pmid..........c8d8300cdac1274e94d6e82c1ed9bb45