Journal: Nature Communications
Article Title: Highly parallel profiling of the activities and specificities of Cas12a variants in human cells
doi: 10.1038/s41467-025-57150-9
Figure Lengend Snippet: a PAM compatibilities of the Cas12a variants in human cells. A heat map showing the average indel frequencies induced by the Cas12a variants at the 15 PAMs that are currently known to be recognized by Cas12a. b Pearson correlation coefficients between the Cas12a variant induced indel frequencies measured at the target sequences containing the same protospacers 7 days after transduction of Cas12a variant lentivirus into libraries integrated HEK293T cells. c Scatter plots showing the correlations of the five cases that are labeled as 1, 2, 3, 4 and 5 in the b . The Spearman correlation coefficient ( ρ ) and the Pearson correlation coefficient ( r ) are shown. In b and c , 15 PAM sequences were used for the analyses. d Comparison of the frequency of indels induced by the Cas12a variants at TTTA, TTTC, TTTG PAM; The boxes represent the 25th, 50th, and 75th percentiles, and the whiskers represent the 10th and 90th percentiles. The numbers of analyzed target sequences ( n ) are as follows: n = 1195, 1223, 1224, 1157, 1191, 1178, 1171 for TTTA, n = 1198, 1642, 1621, 1560, 1591, 1585, 1569 for TTTC, n = 1711, 1753, 1729, 1682, 1706, 1694, 1687 for TTTG (As variants); n = 1240, 1232 for TTTA, n = 1649, 1645, n = 1762, 1775 for TTTG (Eb variants); n = 1295, 1283 for TTTA, n = 1675, 1674, n = 1809, 1805 for TTTG (Lb2 variants); n = 1250 for TTTA, n = 1653 for TTTC, n = 1780 for TTTG (CeCas12a); n = 1202, 1216, 1244, 1300, 1243, 1199, 1210, 1165, 1174 for TTTA, n = 1629, 1623, 1682, 1615, 1646, 1647, 1597, 1596, 1623 for TTTC, n = 1753, 1755, 1809, 1738, 1780, 1778, 1699, 1697, 1748 for TTTG (LbCas12a variants); n = 1205, 1224, 1201 for TTTA, n = 1622, 1632, 1618 for TTTC, n = 1742, 1761, 1745 for TTTG (FnCas12a variants); paired t -test, two tailed. Source data are provided as a Source Data file.
Article Snippet: AsCas12a (Addgene, #69982), LbCas12a (Addgene, #69988), FnCas12a (Addgene, #69976), Lb2Cas12a (Addgene, #69983) human expression plasmids were purchased from the non-profit plasmid repository Addgene.
Techniques: Variant Assay, Transduction, Labeling, Comparison, Two Tailed Test
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